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Anti-Aurora A 抗体 [35C1] (ab13824)

  • Datasheet
  • SDS
Reviews (6)Q&A (9)References (70)

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Western blot - Anti-Aurora A antibody [35C1] (ab13824)
  • Immunocytochemistry - Anti-Aurora A antibody [35C1] (ab13824)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [35C1] (ab13824)
  • Flow Cytometry - Anti-Aurora A antibody [35C1] (ab13824)

Key features and details

  • Mouse monoclonal [35C1] to Aurora A
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG2b

Conjugates logo Related conjugates and formulations

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関連製品

製品の概要

  • 製品名

    Anti-Aurora A antibody [35C1]
    Aurora A 一次抗体 製品一覧
  • 製品の詳細

    Mouse monoclonal [35C1] to Aurora A
  • 由来種

    Mouse
  • アプリケーション

    適用あり: WB, ICC/IF, Flow Cytmore details
  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse
  • 免疫原

    Recombinant full length protein corresponding to Human Aurora A.

  • ポジティブ・コントロール

    • Human HeLa and mouse M-ICc12 cell lysates for Western blotting and human 293 or mouse LLC1 cell lines for IF. Flow Cyt: HeLa cells. ICC: HeLa cells
  • 特記事項

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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

    pH: 7.40
    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

    35C1
  • ミエローマ

    Sp2/0-Ag14
  • アイソタイプ

    IgG2b
  • 研究分野

    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Spindle
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Aurora
    • Cancer
    • Cell cycle
    • Cell division
    • Cancer
    • Tumor biomarkers
    • Other

関連製品

  • Alternative Versions

    • HRP Anti-Aurora A antibody [35C1] (ab199220)
    • PE Anti-Aurora A antibody [35C1] (ab216698)
    • Alexa Fluor® 555 Anti-Aurora A antibody [35C1] (ab217712)
  • 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 IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant human Aurora A protein (ab42595)

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ab13824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
WB (5)
Use a concentration of 1 µg/ml. Detects a band of approximately 46 kDa.
ICC/IF (1)
Use a concentration of 5 µg/ml.
Flow Cyt
Use 2µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

特記事項
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 46 kDa.
ICC/IF
Use a concentration of 5 µg/ml.
Flow Cyt
Use 2µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

ターゲット情報

  • 機能

    Contributes to the regulation of cell cycle progression. Required for normal mitosis. Associates with the centrosome and the spindle microtubules during mitosis and functions in centrosome maturation, spindle assembly, maintenance of spindle bipolarity, centrosome separation and mitotic checkpoint control. Phosphorylates numerous target proteins, including ARHGEF2, BRCA1, KIF2A, NDEL1, PARD3, PLK1 and BORA. Regulates KIF2A tubulin depolymerase activity (By similarity). Required for normal axon formation. Plays a role in microtubule remodeling during neurite extension. Important for microtubule formation and/or stabilization.
  • 組織特異性

    Highly expressed in testis and weakly in skeletal muscle, thymus and spleen. Also highly expressed in colon, ovarian, prostate, neuroblastoma, breast and cervical cancer cell lines.
  • 配列類似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
    Contains 1 protein kinase domain.
  • 翻訳後修飾

    Activated by phosphorylation at Thr-288; this brings about a change in the conformation of the activation segment. Phosphorylation at Thr-288 varies during the cell cycle and is highest during M phase. Autophosphorylated at Thr-288 upon TPX2 binding. Phosphorylated upon DNA damage, probably by ATM or ATR.
    Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by the anaphase-promoting complex (APC), leading to its degradation by the proteasome.
  • 細胞内局在

    Cytoplasm > cytoskeleton > centrosome. Cytoplasm > cytoskeleton > spindle pole. Detected at the neurite hillock in developing neurons (By similarity). Localizes on centrosomes in interphase cells and at each spindle pole in mitosis.
  • Target information above from: UniProt accession O14965 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: 6790 Human
    • Entrez Gene: 20878 Mouse
    • Omim: 603072 Human
    • SwissProt: O14965 Human
    • SwissProt: P97477 Mouse
    • Unigene: 250822 Human
    • Unigene: 249363 Mouse
    • 別名

      • AIK antibody
      • ARK-1 antibody
      • ARK1 antibody
      • AURA antibody
      • Aurka antibody
      • Aurora 2 antibody
      • Aurora A antibody
      • Aurora kinase A antibody
      • Aurora-related kinase 1 antibody
      • Aurora/IPL1 like kinase antibody
      • AURORA/IPL1-like kinase antibody
      • Aurora/IPL1-related kinase 1 antibody
      • AURORA2 antibody
      • Breast tumor-amplified kinase antibody
      • BTAK antibody
      • hARK1 antibody
      • IAK antibody
      • IPL1 related kinase antibody
      • MGC34538 antibody
      • OTTHUMP00000031340 antibody
      • OTTHUMP00000031341 antibody
      • OTTHUMP00000031342 antibody
      • OTTHUMP00000031343 antibody
      • OTTHUMP00000031344 antibody
      • OTTHUMP00000031345 antibody
      • OTTHUMP00000166071 antibody
      • OTTHUMP00000166072 antibody
      • PPP1R47 antibody
      • Protein phosphatase 1, regulatory subunit 47 antibody
      • Serine/threonine kinase 15 antibody
      • Serine/threonine kinase 6 antibody
      • Serine/threonine protein kinase 15 antibody
      • Serine/threonine-protein kinase 15 antibody
      • Serine/threonine-protein kinase 6 antibody
      • Serine/threonine-protein kinase aurora-A antibody
      • STK15 antibody
      • STK6 antibody
      • STK6_HUMAN antibody
      • STK7 antibody
      see all

    画像

    • Western blot - Anti-Aurora A antibody [35C1] (ab13824)
      Western blot - Anti-Aurora A antibody [35C1] (ab13824)
      All lanes : Anti-Aurora A antibody [35C1] (ab13824) at 5 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 46 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?
      Additional bands at: 125 kDa (possible non-specific binding), 55 kDa (possible non-specific binding)


      Exposure time: 20 minutes


      This blot was produced using 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200v for 50 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab13824 over night at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

    • Immunocytochemistry - Anti-Aurora A antibody [35C1] (ab13824)
      Immunocytochemistry - Anti-Aurora A antibody [35C1] (ab13824)

      This data was developed using the same antibody clone in a different buffer formulation without PBS and sodium azide (ab264552)

      ab264552 staining Aurora A in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab264552 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

    • Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [35C1] (ab13824)
      Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [35C1] (ab13824)This image is courtesy of an Abreview submitted by Dr Kirk McManus
      ab13824, at 1/2000 dilution, detecting Aurora A (green) in Hela Cells in conjunction with a Goat anti-mouse secondary antibody conjugated to Cy3®. Cells were fixed with methanol and counterstained with DAPI. Please refer to abreview for further details.

      See Abreview

    • Flow Cytometry - Anti-Aurora A antibody [35C1] (ab13824)
      Flow Cytometry - Anti-Aurora A antibody [35C1] (ab13824)
      Overlay histogram showing HeLa cells stained with ab13824 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13824, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    プロトコール

    • Mouse on Mouse staining 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 (70)

    ab13824 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。

    ab13824 は 70 報の論文で使用されています。

    • Nehlig A  et al. Reciprocal regulation of Aurora kinase A and ATIP3 in the control of metaphase spindle length. Cell Mol Life Sci 78:1765-1779 (2021). PubMed: 32789689
    • Chen X  et al. Aurka loss in CD19+ B cells promotes megakaryocytopoiesis via IL-6/STAT3 signaling-mediated thrombopoietin production. Theranostics 11:4655-4671 (2021). PubMed: 33754019
    • Liu F  et al. Knockdown of AURKA sensitizes the efficacy of radiation in human colorectal cancer. Life Sci 271:119148 (2021). PubMed: 33545203
    • Blazejewski SM  et al. High-throughput kinase inhibitor screening reveals roles for Aurora and Nuak kinases in neurite initiation and dendritic branching. Sci Rep 11:8156 (2021). PubMed: 33854138
    • Xu L  et al. Feedback control of PLK1 by Apolo1 ensures accurate chromosome segregation. Cell Rep 36:109343 (2021). PubMed: 34260926
    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 15 Abreviews or Q&A

    Western blot abreview for Anti-Aurora A antibody [35C1]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Brain prefrontal cortex)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    15 µg
    Specification
    Brain prefrontal cortex
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 14 2016

    Western blot abreview for Anti-Aurora A antibody [35C1]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Huh7 (liver))
    Specification
    Huh7 (liver)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Caroline Gleason

    Verified customer

    投稿 Apr 30 2014

    Western blot abreview for Anti-Aurora A antibody [35C1]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Primary keratinocytes)
    Loading amount
    20 µg
    Specification
    Primary keratinocytes
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 26 2008

    Western blot abreview for Anti-Aurora A antibody [35C1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SW620, IMR-32, 293, MCF7)
    Loading amount
    20 µg
    Specification
    SW620, IMR-32, 293, MCF7
    Gel Running Conditions
    Reduced Denaturing (12.5)
    Blocking step
    5% milk in PBS + 0.1% Tween as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Mr. Antibody Solutions

    Verified customer

    投稿 Jun 24 2008

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Aurora A antibody [35C1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Read More

    DR. Kirk Mcmanus

    Verified customer

    投稿 Apr 19 2007

    Question

    My technician forwarded your note below to me and I simply do not understand your response. You say that the antibody noted below is the best bet as the peptide has a fair amount of alignment between human and Chlamy Aurora A. Well, although the two proteins have 67% homology, there is no overlap at all that I can detect between the human peptide immunogen and any part of the Chlamy Aurora A sequence.

    Have you made a mistake, or posted an incorrect ab link? I am at a loss for understanding your suggested ab (and if I hadn't looked, I would be at a loss for about $*** as well).

    thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 04 2012

    Answer

    Thank you for your reply.

    I apologize for the confusion, there was a difference in the numerical order of amino acids listed in the alignment I was performing. You are correct, ab115883 should not be chosen as the immunogen sequence does not overlap with the Chlamy Aurora A sequence.

    I reran the search using the proper numerical organization and came up with the following antibody:
    ab61114 (https://www.abcam.com/Aurora-A-antibody-ab61114.html): the immunogen sequence is around the human Aurora A amino acid 288. The human and Chlamy sequences are quite homologous in this region, as amino acids 285 and 288 are the only two mismatched amino acids from 275-301.

    I'm actually confident enough in the homology within this region to offer our Abtrial program for testing ab61114 in your Chlamy sequences. Information regarding this program is given below:

    To our knowledge, ab61114 has not been tested in C. reinhardtii. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

    Simply follow these easy steps below to apply for our AbTrial Program:

    1. Reply to this email, letting us know you are interested in testing this product.

    2. Our scientists will email you an inactive personal discount code for the value of the product.

    3. Purchase and test the product at the regular price.

    4. Submit your results, including your discount code in the additional notes section of your Abreview.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    The Terms and Conditions of this offer can be found at: https://www.abcam.com/abtrial

    Additionally, until 12/16/12 we are running a rabbit monoclonal antibody promotion, our "RabMab" promotion, which entitles you to a free rabbit monoclonal antibody with purchase of a primary antibody. I found a rabbit monoclonal antibody ab108353 (https://www.abcam.com/Aurora-A-antibody-EPR5026-ab108353.html) that has 54% homology with the Chlamy sequence and might be worth a try, otherwise you could choose a different Rabbit monoclonal antibody that you desire. Details of the RabMab promotion are given below:
    https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    Read More

    Abcam Scientific Support

    Answered on Dec 05 2012

    Question

    Customer inquiry about Anti-Aurora A antibody for Chlamydomonas reinhardtii. Only 67.2% identity between human and C. reinhardtii sequences. Would like a search for Aurora A antibodies with immunogen that is in a region of increased identity.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 04 2012

    Answer

    Thank you for contacting Abcam.

    We discussed potential anti-Aurora A antibodies to detect Chlamydomonas reinhardtii Aurora A. Since C. reinhardtii Aurora A has 67.2% homology with human Aurora A, this does not qualify for our AbTrial program, but a positive or negative Abreview can be submitted explaining your experience to give you AbPoints which can be used for an amount off of a future purchase or Amazon gift cards.

    I searched through our anti-Aurora A antibodies, and ab115883 is likely your best option should you choose to proceed
    https://www.abcam.com/aurora-a-antibody-ab115883.html

    The immunogen for this antibody is from amino acids 268-277, which has a fair amount of alignment between human and C. reinhardtii Aurora A.
    Human: http://www.uniprot.org/uniprot/O14965
    C. reinhardtii: http://www.uniprot.org/uniprot/A8ISU1

    Additionally, we currently have a RabMab promotion running until 12/16/12, which entitles you to a Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last. Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Dec 04 2012

    Question


    I need help identifying a paper that cites this Aurora A antibody in an immunoprecipitation, please. The reference cited online does not actually use it in an IP:



    Lefebvre C et al. A human B-cell interactome identifies MYB and FOXM1 as master regulators of proliferation in germinal centers. Mol Syst Biol 6:377 (2010). ICC/IF, IP; Human.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 27 2012

    Answer

    A paper regarding use of the antibody in IP is Cremet JY et al., 2003 which is the last one on the
    publication list on our datasheet for ab13824. http://www.ncbi.nlm.nih.gov/pubmed/12619897?dopt=Abstract
    This also appears to be the licensors original publication regarding this clone so there might be some extra information in it that will
    be helpful.

    Read More

    Abcam Scientific Support

    Answered on Sep 27 2012

    Question

    I am doing IHC-P and WB; IHC-P is of mouse brain tumor of injected human tumor cells; WB of tumor cell lysate shows expected band size; IHC shows no staining; IHC used primary at 1:250; secondary is HRP conjugated.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 06 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.

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

    Please note that this 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.

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    Abcam Scientific Support

    Answered on Jul 06 2012

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No staining at all SAMPLE normal mouse preimplantation embryos (2-cell embryo) PRIMARY ANTIBODY Aurora A antibody (ab13824) at 1:200 dilution, RT for 1 h or 4C overnight, DETECTION METHOD confocal microscopy POSITIVE AND NEGATIVE CONTROLS USED stain alpha-tubulin to show spindle. alpha-tubulin got beatiful staining. ANTIBODY STORAGE CONDITIONS -20C FIXATION OF SAMPLE 3.7% PFA at room temp for 1 h PERMEABILIZATION STEP tried both 0.25% and 0.5% triton x-100 at RT for 30 mins and 1h. BLOCKING CONDITIONS 1% BSA in PBS at 4C overnight SECONDARY ANTIBODY Goat anti mouse Alex 633 conjugated, 1: 200 dilution, 1 h at RT or 4C overnight HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? permeabilzation, primary antibody incubation, secondary antibody incubation ADDITIONAL NOTES please see the attached picture. I am sure the antibody does not function. I spent a lot of time on the staining and hope to get the replacement. Thank you.

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    Abcam community

    Verified customer

    Asked on Aug 13 2007

    Answer

    Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab13824. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked at the protocols you used and have a few questions and suggestions that might help you determine the cause of the no staining results. I would therefore appreciate if you can please clarify the following items. Since you are using mouse cell samples, then fixation in 4% PFA should be done not more than 15 minutes, otherwise your epitopes will be masked. Also, we have a customer who previous used this product and mentions that this antibody only recognizes a centrosome associated epitope in methanol-fixed cells and does not recognize anything in a paraformaldehyde-fixed cell. Please see the abreview by Kirk McManus submitted on 19 April 2007 on the datasheet. Can you confirm that the second antibody also managed to produce good results with other primary antibodies? In some cases, the problem may be with the secondary antibody not working properly. I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me with details of your order (purchase order number, batch number, shipping address/purchasing agent information). Also please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you.

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    Abcam Scientific Support

    Answered on Aug 13 2007

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