Overview

  • Product name

    Anti-SKP2 antibody [EPR3305(2)] - BSA and Azide free
    See all SKP2 primary antibodies
  • Description

    Rabbit monoclonal [EPR3305(2)] to SKP2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore 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

  • General notes

    Ab240263 is the carrier-free version of ab183039. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab240263 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab240263 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
WB Use at an assay dependent concentration. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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 [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



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183039).

    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.

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183039).

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

References

ab240263 has not yet been referenced specifically in any publications.

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