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  1. Link

    ncam1-antibody-rnl-1-ab9018.pdf

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Neuroscience Cell Type Marker Neuron marker Growth Cone
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Anti-NCAM1 抗体 [RNL-1] (ab9018)

  • Datasheet
  • SDS
Reviews (4)Q&A (23)References (24)

Product price, shipping and contact information

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Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)
  • Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-NCAM1 antibody [RNL-1] (ab9018)

Key features and details

  • Mouse monoclonal [RNL-1] to NCAM1
  • Suitable for: ICC, IHC-FoFr
  • Reacts with: Rat, Human
  • Isotype: IgG1

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-NCAM1 antibody [CAL53] (ab237708)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-NCAM1 antibody [RNL-1]
    NCAM1 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [RNL-1] to NCAM1
  • 由来種

    Mouse
  • 特異性

    Neural cell adhesion molecule (NCAM). ab9018 reacts with three isoforms of NCAM: NCAM-120, NCAM-140, and NCAM-180.
  • アプリケーション

    適用あり: ICC, IHC-FoFrmore details
    適用なし: Flow Cyt
  • 種交差性

    交差種: Rat, Human
  • 免疫原

    Tissue, cells or virus corresponding to Human NCAM1. This antibody was raised against the small cell lung cancer cell line NCI-H82.
    Database link: P13591

  • ポジティブ・コントロール

    • IHC-P: Mouse brain tissue. IHC-Fr: Rat brain tissue. ICC/IF: Adult-derived human liver stem cells and hepatic stellate cells.
  • 特記事項

    Punctuate cytoplasmic staining is observed with ab9018. Some staining of red blood cells observed.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 精製度

    Protein G purified
  • ポリ/モノ

    モノクローナル
  • クローン名

    RNL-1
  • ミエローマ

    Sp2/0-Ag14
  • アイソタイプ

    IgG1
  • 研究分野

    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Growth Cone
    • Neuroscience
    • Cell Adhesion Proteins
    • ECM Proteins
    • Neuroscience
    • Neurology process
    • Neural Signal Transduction
    • Stem Cells
    • Hematopoietic Progenitors
    • Hematopoietic Stem Cells
    • Human Lineage Negative
    • Neuroscience
    • Development

関連製品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human NCAM1 protein (ab114198)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab9018の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC (1)
1/100 - 1/200.
IHC-FoFr
1/100 - 1/200.
特記事項
ICC
1/100 - 1/200.
IHC-FoFr
1/100 - 1/200.
追加情報
Is unsuitable for Flow Cyt.

ターゲット情報

  • 機能

    This protein is a cell adhesion molecule involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc.
  • 配列類似性

    Contains 2 fibronectin type-III domains.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • 細胞内局在

    Secreted and Cell membrane.
  • Target information above from: UniProt accession P13591 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 4684 Human
    • Entrez Gene: 24586 Rat
    • Omim: 116930 Human
    • SwissProt: P13591 Human
    • SwissProt: P13596 Rat
    • Unigene: 503878 Human
    • Unigene: 11283 Rat
    • 別名

      • antigen MSK39 identified by monoclonal antibody 5.1H11 antibody
      • antigen recognized by monoclonal antibody 5.1H11 antibody
      • CD56 antibody
      • cell adhesion molecule, neural, 1 antibody
      • MSK 39 antibody
      • MSK39 antibody
      • N-CAM-1 antibody
      • NCAM 1 antibody
      • NCAM antibody
      • NCAM C antibody
      • NCAM-1 antibody
      • NCAM1 antibody
      • NCAM1_HUMAN antibody
      • NCAMC antibody
      • Neural cell adhesion molecule 1 antibody
      • Neural cell adhesion molecule NCAM antibody
      • OTTHUMP00000235666 antibody
      see all

    画像

    • Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)
      Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)

      Immunohistochemistry of H82 cells, showing staining for NCAM1 using ab9018 (1/500).

    • Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)
      Immunocytochemistry - Anti-NCAM1 antibody [RNL-1] (ab9018)Image cropped from Berardis et al., Public Library of Science One, 1, e86137, Fig. 3a.; DOI: 10.1371/journal.pone.0086137 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Immunocytochemical analysis of Adult-Derived Human Liver Stem Cells (ADHLSC) in the left image and Hepatic Stellate Cells (HSC) in the right image. ab9018 was used to label NCAM, which is expressed in the HSC. Cells were fixed in 3.5% paraformaldehyde for 15 minutes at room temperature. Blocking was with 3.3% hydrogen peroxide for 3 minutes. Permeabilization with D-PBS and 1% Triton X-100 for 10 minutes. Cells incubated with ab9018 at 1/100 for 1 hour. DAB staining. Counterstaining with Mayer's hematoxylin. 

    • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-NCAM1 antibody [RNL-1] (ab9018)
      Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-NCAM1 antibody [RNL-1] (ab9018)

      ab9018 at a dilution of 1/500, staining NCAM (Alexa 488 secondary at 1/2000) on rat brain tissue (30µm thick coronal sections) in free floating IHC (see protocol link for detailed description). Images showing neuron body and processes: [A] 40x objective and [B] punctate cytoplasmic staining; 20x objective and [C] neuron + processes; x20 objective. No labeling observed following omission of primary antibody. Images coloured in Photoshop.

      NB: We recommend indirect amplification of immunofluorescence for ab9018

      Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

    プロトコール

    • IHC-FoFr protocol

    Click here to view the general protocols

    データシートおよび資料

    • SDS download

    • Datasheet download

      Download

    参考文献 (24)

    ab9018 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab9018 は 24 報の論文で使用されています。

    • Wang J  et al. Low immune index correlates with favorable prognosis but with reduced benefit from chemotherapy in gallbladder cancer. Cancer Sci 111:219-228 (2020). PubMed: 31729088
    • Rao F  et al. Aligned chitosan nanofiber hydrogel grafted with peptides mimicking bioactive brain-derived neurotrophic factor and vascular endothelial growth factor repair long-distance sciatic nerve defects in rats. Theranostics 10:1590-1603 (2020). PubMed: 32042324
    • Xie S  et al. Hyperion imaging system reveals heterogeneous tumor microenvironment of oral squamous cell carcinoma patients at T1N0M0 stage. Ann Transl Med 8:1513 (2020). PubMed: 33313258
    • Feng Q  et al. Integrative proteomics and immunochemistry analysis of the factors in the necrosis and repair in acetaminophen-induced acute liver injury in mice. J Cell Physiol 234:6561-6581 (2019). PubMed: 30417486
    • Li W  et al. Dl-3-n-Butylphthalide Reduces Cognitive Impairment Induced by Chronic Cerebral Hypoperfusion Through GDNF/GFRa1/Ret Signaling Preventing Hippocampal Neuron Apoptosis. Front Cell Neurosci 13:351 (2019). PubMed: 31456664
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 27 Abreviews or Q&A

    Question

    I want to study the expression pattern of Differentiation markers like(ab82259, ab5392, ab9018, ab18258 and ab4648) in both Human neuroblastoma cell line (ICC) and Rat Brain section-Frozen (IHC-Fr) not fixed with any fixative.
    Regarding this experiments, I need your help. mentioned below.
    I want to know that, above mentioned antibodies are suitable for both Human and rat Experiments or not.
    need your suggestion regarding suitable secondary antibody for the above primary antibodies(It is good for me, if its is conjugated with HRP insted of Fluorochrome).
    I need standard protocols for ICC and IHC-Fr for the above mentioned antibodies.
    If you are providing Kit for the above mentioned antibodies, please let me know. That is also good for me.
    I need one more clarification that is the difference between IHC-Fr and IHC-FoFr.
    I am expecting your kind reply as soon as possible.
    Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 19 2013

    Answer


    The antibodies mentioned above are suitable for Rat as well as human samples. ab9018 is yet to be tested in IHC-Fr however as it is tested in IHC-FoFr so it is more likely to work.

    The suitable secondary's are for ab4648, ab82259 and ab9018 are

    https://www.abcam.com/rabbit-polyclonal-secondary-antibody-to-mouse-igg-hl-hrp-ab97046.html

    https://www.abcam.com/goat-polyclonal-secondary-antibody-to-mouse-igg-hl-hrp-ab97023.html

    for ab18258 it is

    https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-hrp-ab6721.html

    for ab5392 it is;

    https://www.abcam.com/goat-polyclonal-secondary-antibody-to-chicken-igy-h-l-hrp-ab97135.html

    The protocols are:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11417

    https://www.abcam.com/index.html?pageconfig=resource&rid=11383

    https://www.abcam.com/index.html?pageconfig=resource&rid=13046

    IHC-Fr is immunohistochemistry on frozen section

    IHC-FoFr is PFA perfusion fixed frozen sections.

    Read More

    Padamjeet Singh

    Abcam Scientific Support

    Answered on Jun 19 2013

    Question

    Many thanks for your email. Sorry for the late reply.

    Yes, it is frustrating about the antibody. I’ve spoken to the our purchaser, and if it is possible for a refund that would be great. We just don’t seem to be getting a signal with the antibody at all.

    Many thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 28 2012

    Answer

    Thank you for contacting us.

    I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for a refund for you.

    Credit note ID: #####

    The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Abcam Scientific Support

    Answered on Aug 28 2012

    Question

    I've have tested the replacement NCAM-1 antibody you kindly sent out last week, but we still haven't had any luck getting a signal by Western blotting or IHC. We used a more sensitive method of diction for the Western and no signal was seen even after 1 hour.

    I have run out of ideas, and I don't think I can try anything else. The fact that no signal is seen with the Western is most worrying as this is our routine method of testing the antibodies.

    Please advise,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 20 2012

    Answer

    Thank you for taking the time to contact me again and provide an update. I am sincerely sorry to hear you have also had difficulty obtaining satisfactory results from the replacement vial of antibody.

    I appreciate your concerns, this is disappointing. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results, so I am not able to suggest anything further. I can reassure you that your enquiries will be kept on our quality monitoring records for our ongoing reference.

    I apologize for the inconvenience. In this case, I suggest it would be appropriate to provide a refund or a credit note for purchase of another product.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed this time.

    Read More

    Abcam Scientific Support

    Answered on Aug 20 2012

    Question

    Many thanks for the NCAM1 that you sent out. We received it today. The only things is that it's the same lot number as before, so I'm not sure if this vial will help. Of course, I will test it first.

    Many thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 10 2012

    Answer

    Thank you for your message.

    I fully understand your concerns and would like to provide some information that I hope will give some reassurance. I can confirmthat we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

    Therefore, I hope the replacement vial should work well. However, please note that the replacement will be covered by the 6 month guarantee should you encounter any problems. I would encourage you to contact me as soon as possible in this case.

    I hope this will be helpful. Please do not hesitate to contact me if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Aug 10 2012

    Question

    Many thanks for your prompt response and for the offer of a replacement vial. A replacement vial would be ideal as I'd really like to use this antibody. I think it would be best to see if I could detect NCAM-1 expression again using a western blot before proceeding with IHC. I’m assuming the rat brain tissue I was using should is a suitable positive control. Just out of interest, are there any other positive controls you could recommend?

    I was hoping that the top bands I could see in the two rat brain tissue lanes in the previously attached western blot may have been NCAM-1. However, as the bands observed were lower than 97kDa suggests that they are too low for NCAM1.

    If you could please let me know when you will ship the replacement, I would be most appreciative.

    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 09 2012

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1143353.

    To check the status of the order please contact our Customer Service team and reference this number.

    With regards to suggests positive controls, I can confirm that rat brain tissue should be a suitable positive control for this protein. There are some images from IHC staining of rat brain on the datasheet, also mouse brain. HT1080 cells woudl also be suitable.

    According to the Unigene EST expression profile, NCAM is expressed in brain, adrenal gland, ear, larynx and lung, and to a lower extent in many other organs:


    http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Hs.503878

    Please note that the free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Aug 09 2012

    Question

    Order Details
    Antibody code: Anti-NCAM antibody [RNL-1] ab9018

    Lot number: GR84221-1

    General Information
    Antibody storage conditions (temperature/reconstitution etc)
    Short term 4C (1-2 weeks). Used most of the antibody during this period as I was trying various antibody dilutions and testing different tissue to see if I could get it to work.

    Description of the problem (high background, low signal, non-specific satining etc.)

    No staining was seen on any slides. I also tried a western blot using mouse muscle (NCAM a marker of muscle satellite cells), rat brain and human cell lines (see attached). No band was seen at the expected molecular weight of 140/180 kDa.

    Sample (Species/Tissue/Cell Type/Cell Line etc.)

    Mouse skeletal muscle tissue (paraffin embedded). Also rat brain as a control (frozen).

    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)Paraffin embedded or frozen sections?
    Paraffin embedded: Paraformaldehyde
    Frozen: Ethanol

    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)

    Tris-EDTA pH8 buffer using microwave technique.

    Has citrate buffer retrieval been tried? What different time points were tried for optimization?
    Citrate buffer hasn’t been tested, but I tried frozen rat brain tissue sections, and once again I could see no signal.

    Permeabilization step


    Blocking conditions (Buffer/time period, Blocking agent etc.)
    Paraffin Embedded Mouse Tissue: Used mouse detection kit to eliminate background staining by the anti-mouse secondary antibody (MaxVision HM01-DS). Used blocking reagents, and 2 ry antibody supplied with kit and followed protocol.
    Frozen Rat Brain Tissue: Used 1xCaesin (1 h 25C).
    Western Blot: 5% Milk in TBS-Tw


    blocking in 1xCaesin, same primary antibodies titrations and detection
    I can recommend to try BSA or serum rather than Caesin to block. Changing blocking agent can sometimes help to improve the results.

    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

    1:50, 1:100 and 1:200 in Dako Diluent (S0809) overnight at 4C.

    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
    Mouse Tissue: 2ry antibody supplied with mouse on mouse kit (manufacturer’s instructions).
    Rat Tissue: 2ry Rabbit antimouse HRP (30 min, 25C). DAKO P0260
    Western Blot: 2ry Goat antimouse HRP (1 h, 25C). Cell Signaling Tech. 7076

    Detection method
    IHC: DAB
    Western Blot: ECL


    Positive and negative controls used (please specify)

    Rat brain tissue that was frozen. Once again, no signal was seen.
    Negative Control: No primary antibody.

    Optimization attempts (problem solving)
    How many times have you tried the IHC?
    5 Times with mouse muscle and rat brain

    Have you run a No Primary control?
    Yes

    Is the current vial of secondary antibody working well with other primary antibodies?
    Yes

    Do you obtain the same results every time?
    Yes


    What steps have you altered?
    Antibody concentration.
    Frozen versus paraffin embedded tissue.
    Rat Brain Control.
    Western Blotting.
    In all case no signal was seen.


    Additional Notes
    Although I wasn't restricted to satellite cells, I was expecting to see clear staining with the rat brain tissue. No signal at the correct molecular weight was seen on a Western Blot.

    We would appreciate if you are also able to provide and image which woudl help us to assess the results
    Image 1: IHC of paraffin embedded mouse skeletal muscle.
    Imange 2: Western blot with NCAM1 (top half) and B-tubulin (bottom half).

    Read More

    Abcam community

    Verified customer

    Asked on Aug 08 2012

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

    I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

    In addition, I can suggest you may like to consider the following suggestions as a check for future experiments:

    1. Antigen retrieval can sometimes require some optimization. I can recommend to try citrate buffer for 2, 5, 10 and 20 minutes, or enzymatic retrieval.

    2. I can recommend to try BSA or serum rather than Caesin to block. Changing blocking agent can sometimes help to improve the results.

    3. Are the current vials of secondary antibody working well with other primary antibodies?

    4. I can suggest to consider Including a permeabilization step which would help to improve the results.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on Aug 08 2012

    Question

    Product code: 9018
    Lot number: GR84221-1

    Inquiry: I have failed to see any positive staining using this antibody on formalin-fixed paraffin-embedded mouse skeletal muscle tissue. (NCAM1 is a marker of muscle satellite cells) and a signal should be seen. I used a mouse on mouse detection kit to eliminate background staining by the anti-mouse secondary antibody (MaxVision HM01-DS). I used a Tris-EDTA pH8 buffer for antigen retrieval and used the reagents with the kit for all other steps. I tested NCAM-1 at 1:50, 1:100 and 1:200 in Dako Diluent (S0809) overnight at 4C. DAB was used for detection and haematoxylin for counterstaining. No staining was seen on any slides. As a positive control, I used rat brain tissue that was frozen and once again, no signal was seen. I used standard IHC procedures for the positive control (blocking in 1xCaesin, same primary antibodies titrations and detection).

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    Abcam community

    Verified customer

    Asked on Aug 06 2012

    Answer

    Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forIHC-P andIHC-FoFr and in mouse and rat samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

    I would appreciate if youare also able to provide an image which would help us to assess the results.

    Thank you for your time and cooperation. We look forward to receiving the completed questionaire.



    Order Details
    Antibody code:

    Lot number:

    Purchase order number
    or preferably Abcam order number:


    General Information
    Antibody storage conditions (temperature/reconstitution etc)


    Description of the problem (high background, low signal, non-specific satining etc.)

    No staining was seen on any slides

    Sample (Species/Tissue/Cell Type/Cell Line etc.)

    mouse skeletal muscle tissue

    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)Paraffin embedded or frozen sections?

    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)

    Tris-EDTA pH8 buffer


    Has citrate buffer retrieval been tried? What different time points were tried for optimization?

    Permeabilization step


    Blocking conditions (Buffer/time period, Blocking agent etc.)

    blocking in 1xCaesin, same primary antibodies titrations and detection

    I can recommend to try BSA or serum rather than Caesin to block. Changing blocking agent can sometimes help to improve the results.

    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

    1:50, 1:100 and 1:200 in Dako Diluent (S0809) overnight at 4C

    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Detection method


    Positive and negative controls used (please specify)

    Rat brain tissue that was frozen and once again, no signal was seen

    Optimization attempts (problem solving)
    How many times have you tried the IHC?



    Have you run a "No Primary" control?
    Yes No


    Is the current vial of secondary antibody working well with other primary antibodies?


    Do you obtain the same results every time?
    Yes No


    What steps have you altered?


    Additional Notes


    We would appreciate if you are also able to provide an image which would help us to assess the results

    Read More

    Abcam Scientific Support

    Answered on Aug 06 2012

    Question

    I am interested in labeling the cell surface of neurons (particularly near the cell body) in live cells for the purpose of identifying neurons in dissociated brains and isolating them by FACS or microdissection for subsequent analysis. Please let me know if you recommend a particular product for this purpose.

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    Abcam community

    Verified customer

    Asked on Jun 28 2012

    Answer

    Thank you for contacting us.

    I suggest having a look at some of the references for our NCAM antibodies such as ab9018, to see if this marker will be appropriate for what you are trying to do. It is not clear to me if NCAM is on all neurons or just a subset, though.

    Click here (or use the following: https://www.abcam.com/NCAM1-antibody-RNL-1-ab9018.html).

    All other markers that I know of are intracellular.

    While searching for marker candidates, I came across a paper which appears to have a good discussion of tissue dissociation techniques,

    Stem Cells. 2007 Jun;25(6):1560-70.

    Please do not hesitate to contact us if you need any more advice or information.

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    Abcam Scientific Support

    Answered on Jun 28 2012

    Question

    I am using this antibody in IHC staining by not seeing any signal.

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    Abcam community

    Verified customer

    Asked on Dec 09 2011

    Answer

    Thank you for contacting us. I am sorry that this antibody did not perform as stated on the datasheet. The above amount will be credited back to your credit card. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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    Abcam Scientific Support

    Answered on Dec 09 2011

    Question

    Thanks for your quick reply. According your protocol, I prepared some slides (attached). I am not very sure whether they are real positive staining. I blocked the slides for 1 hour. But I don’t know the exact concentration of the blocking buffer. Cause I used VECTOR anti-mouse secondary antibody kit (I did staining on mouse tissue).  

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    Abcam community

    Verified customer

    Asked on Dec 05 2011

    Answer

    Thank you for sending those images. Comparing the images you sent and the images from the Allen Institute for Brain Science, see link below: http://mouse.brain-map.org/gene/show/17734 It would appear that your staining is similar to that theirs, maybe a little stronger but it all seems to be good (as long as I am reading you slides correctly and you were looking at the Cerebellum). To try and reduce the intensity of the staining, you could lower the primary antibody concentration to 1/150 and also block for longer. Please let me know if there is anything else I can help you with.    

    Read More

    Abcam Scientific Support

    Answered on Dec 05 2011

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