Anti-SKP2 antibody (ab68455)

Reviews (1)Q&A (3)References (9)

Overview

  • Product name

    Anti-SKP2 antibody
    See all SKP2 primary antibodies

  • Description

    Rabbit polyclonal to SKP2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, ICCmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to Human SKP2 aa 2-20 (N terminal). Different from the related rat and mouse sequences by three amino acids.
    Sequence:

    HRKHLQEIPDLSSNVATSF


    Database link: Q13309

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

Applications

Our Abpromise guarantee covers the use of ab68455 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
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 48 kDa.
IHC-P Use a concentration of 1 - 2 µg/ml.
ICC Use a concentration of 1 - 2 µg/ml. Acetone fixed cells. Antigen retrieval by Pepsin and Trypsin is required.

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

    see all

  • 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

  • Western blot - Anti-SKP2 antibody (ab68455)
    Western blot - Anti-SKP2 antibody (ab68455)
    All lanes : Anti-SKP2 antibody (ab68455) at 1 µg/ml

    Lane 1 : Whole cell lysates prepared from Hela cells
    Lane 2 : Whole cell lysates prepared from 6T-CEM cells

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : HRP conjugated Goat anti-rabbit IgG at 1/3000 dilution

    Predicted band size: 48 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody (ab68455)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody (ab68455)

    ab68455 at 1µg/ml staining SKP2 in Human mammary cancer tissue sections by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). The tissue underwent enzymatic antigen retrieval in pepsin and trypsin. A Biotin-conjugated Goat anti-rabbit IgG was used as secondary at 1/200 dilution.

  • Western blot - Anti-SKP2 antibody (ab68455)
    Western blot - Anti-SKP2 antibody (ab68455)
    All lanes : Anti-SKP2 antibody (ab68455) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : Human placenta tissue lysate - total protein (ab29745)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 53 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 29 kDa, 38 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute


    The predicted molecular weight of SKP2 is 48 kDa (SwissProt), however we expect to observe a banding pattern at 53 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • Immunoprecipitation - Anti-SKP2 antibody (ab68455)
    Immunoprecipitation - Anti-SKP2 antibody (ab68455)
    SKP2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to SKP2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab68455.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 53kDa; SKP2

References

This product has been referenced in:

  • Yang Y  et al. Skp2 inhibitor SKPin C1 decreased viability and proliferation of multiple myeloma cells and induced apoptosis. Braz J Med Biol Res 52:e8412 (2019). Read more (PubMed: 31038581) »
  • Yan W  et al. Inhibition of SKP2 Activity Impaired ATM-Mediated DNA Repair and Enhanced Sensitivity of Cisplatin-Resistant Mantle Cell Lymphoma Cells. Cancer Biother Radiopharm N/A:N/A (2019). Read more (PubMed: 31025879) »
See all 9 Publications for this product

Publishing research using ab68455? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-SKP2 antibody

 Inconclusive
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (Amersham ECL Gel 8-16%)
Sample
Chinese Hamster Cell lysate - whole cell (CHO DG44 cell line)
Specification
CHO DG44 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Apr 16 2014

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


thanks for your reply. After consulting with people in the lab we were wondering if you could send us a sample of both your antibodies so that we can test in our cell model.
Let me know if this is possible. In case we have to pick one is there a way you can tell us which one most likely would recognize the murine form?
Thanks again

Read More
Answer

Thank you for your reply.

We do not have small test samples available to send.

I would say both ab19877 and ab68887 have an equal likelihood of reacting with murine samples as both have similar percentage identity to the immunogens used to produce those antibodies.

Please let me know which antibody you would like and I'll make sure to send it out right away.

Read More

Question

.
I am writing in reference of the skp2 antibody ab68544 that we have recently purchased by you with .
We were unable to detect a specific band in protein lysates from mouse smooth muscle cells, our primary cell model. Therefore we tested in MEFs wild type versus MEFs Skp2 null (negative control), see attachment.
As you can see there are no differences.

Could we exchange with another skp2 antibody that works successfully in Western Blot analysis of mouse cell protein extracts?

Read More
Answer

Thank you for your enquiry and for including the western blotting data.

I am sorry to see this vial of ab68455 is not working as expected. I would be happy to send you another vial of a different anti-SKP2 antibody. At this time none of our other SKP2 antibodies have been tested for reactivity to mouse SKP2.

However, two antibodies to this protein that have a good chance of cross reacting with mouse SKP2, based on comparison of immunogen sequence with that of mouse, are ab19877 and ab68887. I would be happy to send either of these antibodies to you free of charge to test in your system.

I look forward to hearing from you.

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