Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SKP2 antibody [EPR3305(2)] (ab183039)

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Overview

  • Product name

    Anti-SKP2 antibody [EPR3305(2)]
    See all SKP2 primary antibodies

  • Description

    Rabbit monoclonal [EPR3305(2)] to SKP2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human SKP2 aa 1-100. The exact sequence is proprietary.
    Database link: Q13309

  • Positive control

    • WB: C6, NIH/3T3, F9, A549, MDA-MB-231, Jurkat, MCF7, HepG2, and NCCIT cell lysates. ICC: 293T cells, FC: HeLa cells, IHC-P: Human kidney clear cell carcinoma tissue

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab183039 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 1/100.
Flow Cyt 1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/200. Detects a band of approximately 48 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

    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 [EPR3305(2)] (ab183039)
    Western blot - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    All lanes : Anti-SKP2 antibody [EPR3305(2)] (ab183039) at 1/200 dilution

    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 - ab183039 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab183039 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. Ab183039 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/200 dilution 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody [EPR3305(2)] (ab183039)

    Immunofluorescenct analysis of 4% paraformaldehyde fixed 293T cells labeling SKP2 with ab183039 at 1/100 followed by Goat anti rabbit IgG(Alexa Fluor® 488) at 1/200 (green). Cells were counter stained with Dapi (blue).

  • Flow Cytometry - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    Flow Cytometry - Anti-SKP2 antibody [EPR3305(2)] (ab183039)

    Flow cytometric analysis of 2% paraformaldehyde fixed HeLa cells labeling SKP2 with ab183039 at 1/70 followed by Goat anti rabbit IgG (FITC) at 1/150. Rabbit monoclonal IgG was used as Isotype control.

  • Western blot - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    Western blot - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    All lanes : Anti-SKP2 antibody [EPR3305(2)] (ab183039) at 1/200 dilution

    Lane 1 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates with 5% NFDM/TBST
    Lane 5 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 6 : A549 (Human lung carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 7 : F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 8 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates with 5% NFDM/TBST
    Lane 9 : C6 (Rat glial tumor glial cell) whole cell lysate with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 48 kDa


    Exposure time: 180 seconds


    We recommend to increase the amount of samples or decrease antibody dilution to get clear bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody [EPR3305(2)] (ab183039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SKP2 antibody [EPR3305(2)] (ab183039)

    immunohistochemical analysis of paraffin embedded Human kidney clear cell carcinoma tissue labeling SKP2 with ab183039 at 1/50 followed by secondary staining with Ready to use HRP Polymer for Rabbit IgG and counterstained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Song J  et al. Rottlerin promotes autophagy and apoptosis in gastric cancer cell lines. Mol Med Rep 18:2905-2913 (2018). Read more (PubMed: 30015872) »
  • Dai Y  et al. Activation of anaphase-promoting complex by p53 induces a state of dormancy in cancer cells against chemotherapeutic stress. Oncotarget 7:25478-92 (2016). WB . Read more (PubMed: 27009858) »
See all 2 Publications for this product

Publishing research using ab183039? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-SKP2 antibody [EPR3305(2)]

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde
Read More

Dr. Kirk Mcmanus

Verified customer

Submitted Nov 07 2017

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