Overview

  • Product name
  • Description
    Rabbit polyclonal to SKP2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Dog
  • Immunogen

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

    Read Abcam's proprietary immunogen policy (Peptide available as ab26153.)

  • Positive control
    • 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

Properties

Applications

Our Abpromise guarantee covers the use of ab19877 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
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).

Target

  • Function
    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.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Contains 1 F-box domain.
    Contains 9 LRR (leucine-rich) repeats.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • 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.
  • ICC/IF image of ab19877 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab19877 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • 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). Read more (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 . Read more (PubMed: 19648963) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

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
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Sep 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Mr. Antibody Solutions

Verified customer

Submitted Feb 11 2009

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.

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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,

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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