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遺伝子ノックアウト細胞株 検証済

Anti-SKP2 抗体 (ab19877)

Reviews (2)Q&A (4)References (8)
Western blot - Anti-SKP2 antibody (ab19877)
  • Western blot - Anti-SKP2 antibody (ab19877)
  • Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody (ab19877)

Key features and details

  • Rabbit polyclonal to SKP2
  • Suitable for: ICC/IF, WB
  • Knockout validated
  • Reacts with: 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-SKP2 antibody [EPR3305(2)] (ab183039)
  • 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-SKP2 antibody
    SKP2 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to SKP2

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: ICC/IF, WBmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Dog

  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human SKP2.

    Immunogen の所有権に関して (Peptide available as ab26153.)

  • ポジティブ･コントロール

    • Recombinant Human SKP2 protein (ab114385) can be used as a positive control in WB. Human heart tissue lysate This antibody gave a positive signal in the following formaldehyde fixed cell line: HeLa

  • 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ab19877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/IF
Use a concentration of 5 µg/ml.
WB (2)
Use a concentration of 1 µg/ml. Detects a band of approximately 46 kDa (predicted molecular weight: 48 kDa).
特記事項
ICC/IF
Use a concentration of 5 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 46 kDa (predicted molecular weight: 48 kDa).

ターゲット情報

  • 機能

    Substrate recognition component of a SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins involved in cell cycle progression, signal transduction and transcription. Specifically recognizes phosphorylated CDKN1B/p27kip and is involved in regulation of G1/S transition. Degradation of CDKN1B/p27kip also requires CKS1. Recognizes target proteins ORC1, CDT1, RBL2, MLL, CDK9, RAG2, FOXO1A, UBP43, and probably MYC, TOB1 and TAL1. Degradation of TAL1 also requires STUB1. Recognizes CDKN1A in association with CCNE1 or CCNE2 and CDK2.

  • パスウェイ

    Protein modification; protein ubiquitination.

  • 配列類似性

    Contains 1 F-box domain.
    Contains 9 LRR (leucine-rich) repeats.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CDK2/Cyclin A associated protein p45 antibody
      • Cyclin A/CDK2 associated protein p45 antibody
      • Cyclin-A/CDK2-associated protein p45 antibody
      • F box protein Skp2 antibody
      • F box/LRR repeat protein 1 antibody
      • F-box protein Skp2 antibody
      • F-box/LRR-repeat protein 1 antibody
      • FBL 1 antibody
      • FBL1 antibody
      • FBXL 1 antibody
      • FBXL1 antibody
      • FLB 1 antibody
      • FLB1 antibody
      • MGC1366 antibody
      • p45 antibody
      • p45skp2 antibody
      • S phase kinase associated protein 2 (p45) antibody
      • S phase kinase associated protein 2 antibody
      • S-phase kinase-associated protein 2 antibody
      • S-phase kinase-associated protein 2 E3 ubiquitin protein ligase antibody
      • SKP 2 antibody
      • Skp2 antibody
      • SKP2_HUMAN antibody
      see all

    画像

    • Western blot - Anti-SKP2 antibody (ab19877)
      Western blot - Anti-SKP2 antibody (ab19877)
      All lanes : Anti-SKP2 antibody (ab19877) at 1 µg/ml

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : SKP2 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : MCF7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 48 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab19877 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab19877 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in SKP2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SKP2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab19877 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-SKP2 antibody (ab19877)
      Western blot - Anti-SKP2 antibody (ab19877)
      All lanes : Anti-SKP2 antibody (ab19877) at 1 µg/ml

      Lane 1 : Human heart tissue lysate
      Lane 2 : Human brain tissue lysate
      Lane 3 : Human kidney normal tissue lysate
      Lane 4 : Human testis tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 48 kDa
      Observed band size: 46 kDa why is the actual band size different from the predicted?



      ab19877 detects a band at approximately 46 kDa that corresponds in size to that predicted for SKP2 (48 kDa). This band is detected in several tissue lysates. This antibody was tested in normal liver tissue lysates, but no band was detected.
    • Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody (ab19877)
      Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody (ab19877)

      ab19877 stained in HeLa cells. Cells were fixed with 4% paraformaldehyde (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab19877 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

    参考文献 (8)

    ab19877 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。

    ab19877 は 8 報の論文で使用されています。

    • Zhang C  et al. Isoalantolactone inhibits pancreatic cancer proliferation by regulation of PI3K and Wnt signal pathway. PLoS One 16:e0247752 (2021). PubMed: 33661942
    • Yang Y  et al. A non-canonical monovalent zinc finger stabilizes the integration of Cfp1 into the H3K4 methyltransferase complex COMPASS. Nucleic Acids Res 48:421-431 (2020). PubMed: 31724694
    • Alza L  et al. FAK Inhibition Induces Glioblastoma Cell Senescence-Like State through p62 and p27. Cancers (Basel) 12:N/A (2020). PubMed: 32349327
    • Liu L  et al. PTEN inhibits non-small cell lung cancer cell growth by promoting G0/G1 arrest and cell apoptosis. Oncol Lett 17:1333-1340 (2019). PubMed: 30655903
    • Wang L  et al. Mechanism of Low Expression of miR-30a-5p on Epithelial-Mesenchymal Transition and Metastasis in Ovarian Cancer. DNA Cell Biol 38:341-351 (2019). PubMed: 30839226
    • Zhuang K  et al. Gastrin induces multidrug resistance via the degradation of p27Kip1 in the gastric carcinoma cell line SGC7901. Int J Oncol 50:2091-2100 (2017). PubMed: 28498440
    • Lo Sasso G  et al. Liver X receptors inhibit proliferation of human colorectal cancer cells and growth of intestinal tumors in mice. Gastroenterology 144:1497-507, 1507.e1-13 (2013). PubMed: 23419360
    • Kelly-Spratt KS  et al. Inhibition of PI-3K restores nuclear p27Kip1 expression in a mouse model of Kras-driven lung cancer. Oncogene 28:3652-62 (2009). WB . PubMed: 19648963

    レビューと Q&A

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    レビューを送る 質問を送る

    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-SKP2 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U87MG tumour cell line)
    Loading amount
    50 µg
    Specification
    U87MG tumour cell line
    Treatment
    U87MG cells treated with LY294002 for 6h
    Gel Running Conditions
    Reduced Denaturing (10% tris glycine)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    投稿 Sep 22 2010

    Western blot abreview for Anti-SKP2 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela HEK293 MCF7)
    Loading amount
    20 µg
    Specification
    Hela HEK293 MCF7
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Mr. Antibody Solutions

    Verified customer

    投稿 Feb 11 2009

    Question


    we would like to test in WB the polycolonal ab19877.
    If you could send us a vial here is the address:

    Read More
    Answer

    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 for one vial of ab19877.

    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.

    Read More

    Question

    Thank you for your email regarding the Skp2 antibody. I appologise if my last email was slightly strongly worded. I accept that changing blocking reagents and time etc can sometime effect the quality of westerns but in my experience these sorts of changes rarely improve a western that has no reactivity to one that is presentable. In this case I don't think it is going to be very fruitful spending a lot of time trouble shooting this western. Thank you for your offer of a refund for the antibody. As we are a charity funded lab every £200 pound for an antibody that does not produce result makes a big difference to us. I am not sure how the refund procedure works. I you need to send a credit note you may need to contact our departmental administrator, Mrs Dawn Wallace at email D.T.Wallace@bristol.ac.uk Thank you again for your help with this.

    Read More
    Answer

    Thank you for your reply. I have asked our accounts department to arrange this credit note to pay for the order. I hope this unfortunate matter will not deter you from purchasing at Abcam in the future. Please be assured that I will closely monitor the quality of this antibody following your complaint.

    Read More

    Question

    To clarify, the conditions I used (which were identical to those used for the [anoter compagny] antibody, appart from primary Ab dilution/concentration) 100ug total cell lysate/lane protein transferred to PVDF membrane membrane blocked in 6% milk powder in TBS+0.1% tween20 for 7 hours at room temp. Primary antibody diluted in TBS+0.1% tween 20 at 1ug/ml Blot incubated in antibody overnight at 4oC Blot repeatedly washed in large volumes of TBS+0.1% tween20 2nd Ab = 1:3000 [anoter compagny] swine anti-rabbit-HRP Blot repeated washed in large volumes of TBS-T Previous blots detected with standard ECL - no signal therefore the blot I sent you was detected with ECL plus. thanks for your help

    Read More
    Answer

    Thank you very much for your reply and detailed protocol which is very similar to the one we recommend. In our experience too much blocking can prevent the antibody from binding adequately to the membrane unfortunately. I would therefore recommend to block the membrane for 1 hour only and as you have done in the past to incubate the primary antibody in TBST only. We also have very good results with 5% BSA in the same conditions. If you still have problems with a reduced blocking time please let me know by e-mail or do not hesitate to call me and I will arrange a refund on your order immediately.

    Read More

    Question

    Dear Abcam, I recently purchased your rabbit anti Skp2 antibody (ab19877). I have now tested this antibody on western blots containing rat, human and mouse cell lysates and compared it to [another company] anti Skp2 antibody using the sample samples run at the same time. I would like to complain that your antibody does not recognise Skp2 in rat, mouse or human cell lysates. As you can see from the scans of the Abcam and [another company] blots, Skp2 should be absent in serum deprived cells, strongly induced in FCS stimulated cells and present in asynchronously proliferating cells. This is clearly evident in the strong Skp2 band int he [another company] blot but not at all on the blot using the Abcam antibody. The band in the Abcam blot running at slightly high than the 45k marker is a non-specific band. I would therefore like to request a refund on this antibody as it clearly is not suitable for use in detecting Skp2 on western blots with human or mouse cell lysates. My order number when I originally placed this order was BHI-12057 and the invoice number was 1029721. Thank you for your help,

    Read More
    Answer

    I'm very sorry to hear you are experiencing problems with ab19877. I take your comments very seriously and would appreciate if you could please clarify the conditions you have used the antibody in. It is clear that your samples express high levels of SKP2, therefore there is no doubt that the problem is due to either the quality of the antibody or the conditions it has been used. To eliminate the second possibility as a reason can you please clarify: -what blocking agent was used -what antibody dilution buffer was used -how long the antibody was incubated for -what dilutions were tested Please be assured that should the problem be due to the quality of the product I can certainly arrange for a credit note to be issued to pay for the order 156940 (BHI-12057). I apologise for the delay in resolving this matter and thank you in advance for providing the above information needed,

    Read More

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