Anti-KIF4A/KIF4 抗体 (ab3815)

Reviews (5)Q&A (6)References (6)
Western blot - Anti-KIF4A/KIF4 antibody (ab3815)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF4A/KIF4 antibody (ab3815)

Key features and details

  • Goat polyclonal to KIF4A/KIF4
  • Suitable for: WB, ICC
  • Reacts with: Mouse, Human
  • Isotype: IgG

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-KIF4A/KIF4 antibody [EPR5459] (ab124903)
  • 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-KIF4A/KIF4 antibody
    KIF4A/KIF4 一次抗体 製品一覧

  • 製品の詳細

    Goat polyclonal to KIF4A/KIF4

  • 由来種

    Goat

  • アプリケーション

    適用あり: WB, ICCmore details

  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Rat, Cow, Pig

  • 免疫原

    Synthetic peptide corresponding to Human KIF4A/KIF4 aa 1219-1232 (C terminal).
    Sequence:

    SFFSGCSPIEEEAH


    (Peptide available as ab22955)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • WB: NIH/3T3 nuclear lysate. ICC: U-2 OS cells.

  • 特記事項

    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

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ab3815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 (1)
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 140 kDa.
ICC
Use a concentration of 10 µg/ml.
特記事項
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 140 kDa.
ICC
Use a concentration of 10 µg/ml.

ターゲット情報

  • 機能

    Motor protein that translocates PRC1 to the plus ends of interdigitating spindle microtubules during the metaphase to anaphase transition, an essential step for the formation of an organized central spindle midzone and midbody and for successful cytokinesis. May play a role in mitotic chromosomal positioning and bipolar spindle stabilization.

  • 組織特異性

    Highly expressed in hematopoietic tissues, fetal liver, spleen, thymus and adult thymus and bone marrow. Lower levels are found in heart, testis, kidney, colon and lung.

  • 配列類似性

    Belongs to the kinesin-like protein family. Chromokinesin subfamily.
    Contains 1 kinesin-motor domain.

  • 細胞内局在

    Nucleus matrix. Cytoplasm > cytoskeleton > spindle. Midbody. Chromosome. Not present in the nucleolus. In early mitosis, associated with the mitotic spindle, in anaphase, localized to the spindle midzone and, in telophase and cytokinesis, to the midbody. In late cytokinesis, found in the center of the midbody. Associated with chromosomes at all stages of mitosis.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Chromokinesin antibody
      • Chromokinesin-A antibody
      • Chromosome associated kinesin KIF4A antibody
      • Chromosome-associated kinesin KIF4A antibody
      • FLJ12530 antibody
      • FLJ12655 antibody
      • FLJ14204 antibody
      • FLJ20631 antibody
      • HSA271784 antibody
      • KIF4 antibody
      • KIF4 G1 antibody
      • KIF4A antibody
      • KIF4A_HUMAN antibody
      • KIF4G1 antibody
      • Kinesin family member 4A antibody
      see all

    画像

    • Western blot - Anti-KIF4A/KIF4 antibody (ab3815)
      Western blot - Anti-KIF4A/KIF4 antibody (ab3815)
      All lanes : Anti-KIF4A/KIF4 antibody (ab3815) at 2 µg/ml

      Lane 1 : NIH/3T3 nuclear lysate
      Lane 2 : Human ovary lysate

      Lysates/proteins at 35 µg per lane.

      Predicted band size: 140 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF4A/KIF4 antibody (ab3815)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF4A/KIF4 antibody (ab3815)

      Immunocytochemical analysis of paraformaldehyde-fixed U-2 OS cells permeabilized with 0.15% Triton. Cells were incubated with ab3815 at 10 ug/ml for 1 hour followed by an Alexa Fluor® 488 secondary at 4 ug/ml (green). The nuclear stain is DAPI (blue). Negative control is unimmunized goat IgG at 10 ug/ml, followed by an Alexa Fluor® 488 secondary at 4 ug/ml (green).

    参考文献 (6)

    ab3815 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。

    ab3815 は 6 報の論文で使用されています。

    • Champion L  et al. Dissociation of membrane-chromatin contacts is required for proper chromosome segregation in mitosis. Mol Biol Cell 30:427-440 (2019). PubMed: 30586323
    • Lopez-Camacho C  et al. Core binding factor ß (CBFß) is retained in the midbody during cytokinesis. J Cell Physiol 229:1466-74 (2014). PubMed: 24648201
    • Hu CK  et al. Midbody assembly and its regulation during cytokinesis. Mol Biol Cell 23:1024-34 (2012). ICC/IF . PubMed: 22278743
    • Hu CK  et al. Plk1 negatively regulates PRC1 to prevent premature midzone formation before cytokinesis. Mol Biol Cell 23:2702-11 (2012). ICC/IF ; Human . PubMed: 22621898
    • Hu CK  et al. KIF4 regulates midzone length during cytokinesis. Curr Biol 21:815-24 (2011). PubMed: 21565503
    • Wilmeth LJ  et al. Mutual dependence of Mob1 and the chromosomal passenger complex for localization during mitosis. Mol Biol Cell 21:380-92 (2010). Human . PubMed: 19955215

    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 11 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-KIF4A/KIF4 antibody

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (retina)
    Permeabilization
    Yes - triton-X100
    Specification
    retina
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 28 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KIF4A/KIF4 antibody

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cell line)
    Permeabilization
    Yes - triton .15
    Specification
    HeLa cell line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 09 2012

    Immunoprecipitation abreview for Anti-KIF4A/KIF4 antibody

     Good
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Total protein in input
    10 µg
    Treatment
    500nM nocodazole for 12 hrs
    Immuno-precipitation step
    Protein A/G
    Specification
    HeLa cells
    Read More

    Abcam user community

    Verified customer

    投稿 May 28 2009

    Western blot abreview for Anti-KIF4A/KIF4 antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Treatment
    Nocodazole synchronized for 12hrs
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 23 2009

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KIF4A antibody

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 03 2008

    Question

    Thank you for all your help
    Unfortunately the replacement did not work as well
    It looks that it was a bad lot, rather than a bad vial.
    Pleas advise
    Best Regards

    Read More
    Answer

    Thank you for taking the time tocontact us. I am sorry to hearthe customer has had difficulty obtaining satisfactory results from this second vial of antibody.

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

    I appreciate the time the customer has spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there would be no further tips to provide on this occasion to help improve the results andI apologize for the inconvenience. I'm pleased to offer you acredit note or alternative replacementin compensation, which may be more appropriate in this case than another vial of the same antibody.

    In addition, I would appreciate if you could confirm the following information which will help with our quality monitoring:

    1. Please confirm if the same type of samples have been used in this case (HeLa cells).

    2. Has the same procedure (with the suggestions previously provided) been used?

    3. What is the problem with the results? Is there high background in WB and ICC again?

    4. Has the customer tried fresh samples?

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

    Read More

    Question

    After consulting with the customer she
    asked to receive a new vial
    Thanks
    Best regards

    Read More
    Answer

    Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

    As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab3815 has been added to your order number [BO] 88841. This has Abcam order reference 1112739.

    I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

    Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

    Read More

    Question

    The order details
    Po: ####
    Lot: GR86868
    Invoice: #####
    The customer would like to try different lot
    Please advise

    Read More
    Answer

    Thank you for your reply.

    I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that 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 suggestthe customer has received a bad vial on this occasion.

    Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand the customers concerns and if they prefer a credit note in this case, I will be pleased to arrange this for them.

    I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    Please see to the customer answer at the attached file

    Best regards

    Read More
    Answer

    Thank you for your message and for kindlyproviding this further information.

    I am sorry to hear the suggestions made have not improved the results on this occasion. I would like to reassure you that 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.

    I appreciate the time you have spent on these experiments, and would like to offer a refund or credit note in compensation (providing the product has been purchased in the last 6 months). In order to arrange this, I would appreciate if you could confirm your order number and date of purchase as previously requested?

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

    Read More

    Question

    Product code: 3815
    Lot number: GR86868-1

    Inquiry: Antibody code: ab3815
    Batch number: GR86868-1
    General Information:
    Antibody storage conditions (temperature/reconstitution etc) -20deg
    Description of the problem (high background, low signal, non-specific staining etc.) non-specific staining, no relevant signal
    Sample (Species/Tissue/Cell Type/Cell Line etc.) HELA cells
    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
    Tried both Methanol/Acetone/ or Paraformaldehyde
    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
    Permeabilization step Aceton for Methanol, Triton for formaldehyde
    Blocking conditions (Buffer/time period, Blocking agent etc.) 5% BSA, 1h
    Primary Antibody (Diluent/Dilution/Incubation time, Wash step) 1h RT 1:100, 3 washes PBST
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
    Wash step) Alexa anti goat 594, 1:200, 1h RT
    Detection method Observer. Z1 Microscope (Zeiss)
    Positive and negative controls used (please specify) negative control – no primary Optimization attempts (problem solving):
    How many times have you tried the IHC? 2 times
    Have you run a "No Primary" control? yes Do you obtain the same results every time? yes
    What steps have you altered?
    Additional Notes
    Document Attachment: images of the results may be very helpful. If you wish to add it to the complaint form, please attach them to the reply e-mail.

    Antibody code: ab3815 Batch number: GR86868-1
    Antibody storage conditions (temperature/reconstitution etc) -20
    Description of the problem (high background, wrong band size, more bands, no band etc.) no relevant band, many non-specific bands
    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) HELA cell extract, 293-T cell extract, HELA and 293-T synchronized with Nocodazole for 12h lysates
    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) RIPA buffer + protease inhibitors (merk)
    Amount of protein loaded 30microg
    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) reducing gel 10%
    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) 1h 20min semi-dry transfer, TGB +20% MetOH transfer buffer, blocking with 5% milk
    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) 1:100 in 1% BSA solution, 1h RT or ON 4deg, 6 washes with PBST
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Anti goat 1:5000 (Jackson), 1% BSA, 1h RT, 6 washes with PBST
    Detection method (ECL, ECLPlus etc.) ECL Positive and negative controls used (please specify) OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
    How many times have you tried the Western? 3 times Have you run a "No Primary" control?
    Do you obtain the same results every time? e.g. are the background bands always in the same place? yes
    What steps have you altered? ON incubation with ab, cell syncronization Additional Notes

    Read More
    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 enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab3815 is tested and covered by our 6 month guarantee for use in WB, ICC-IF and inhuman samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if youare able to confirm some further details:

    ICC-IF

    1. Has the quality of the sample been assessed, for example what was the% confluency before staining, and was a spare sample checked for viability using trypan blue count before staining?

    2. Fixation: To prevent over fixation, whichever method is used, I can suggest this should be for 10 minutes only if this has not already been tried. PFA should be 4% only.

    3. Please provide further information regarding the permeabilization. I would suggest to try 0.2% triton, 10 minutes only.

    4. I can recommend to consider reducing the concentration to 1:500 to reduce the background signal.

    5. I would appreciate if you are able to provide an image which would help us to assess the results.

    WB

    1. I can suggest to try reducingthe concentration to 1:500 to reduce the background signal. Incubate overnight 4oC.

    2. Could you confirm if the quality of sample been assessed using a loading control?

    3. Have the samples reduced and denatured?

    4. I would appreciate if you are able to provide an image which would help us to assess the results.

    General:

    1. Please confirm the order number and date of purchase.

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

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    1-10 of 11 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com