Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Cdk5 antibody [EP715Y] - BSA and Azide free (ab220214)

Overview

  • Product name

    Anti-Cdk5 antibody [EP715Y] - BSA and Azide free
    See all Cdk5 primary antibodies
  • Description

    Rabbit monoclonal [EP715Y] to Cdk5 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC, Flow Cyt, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Chicken, Human
  • Immunogen

    Synthetic peptide corresponding to residues in the C-terminus of human Cdk5

  • Positive control

    • WB: HeLa cell lysate. IHC-P: Human brain tissue. Flow Cyt: HeLa cells.
  • General notes

    Ab220214 is the carrier-free version of ab40773. 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.

    ab220214 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 ab220214 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 33 kDa (predicted molecular weight: 33 kDa).
IP 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.
ICC Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

IHC-Fr Use at an assay dependent concentration.

Target

  • Function

    Probably involved in the control of the cell cycle. Interacts with D1 and D3-type G1 cyclins. Can phosphorylate histone H1, tau, MAP2 and NF-H and NF-M. Also interacts with p35 which activates the kinase.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
    Contains 1 protein kinase domain.
  • Cellular localization

    Cytoplasm. Cell projection > lamellipodium. Cell projection > growth cone. In axonal growth cone with extension to the peripheral lamellipodia.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cdk 5 antibody
    • Cdk5 antibody
    • CDK5_HUMAN antibody
    • Cell division protein kinase 5 antibody
    • Crk6 antibody
    • Cyclin dependent kinase 5 antibody
    • Cyclin-dependent kinase 5 antibody
    • Protein kinase CDK5 splicing antibody
    • PSSALRE antibody
    • Serine threonine protein kinase PSSALRE antibody
    • Serine/threonine-protein kinase PSSALRE antibody
    • Tau protein kinase II catalytic subunit antibody
    • TPKII catalytic subunit antibody
    see all

Images

  • All lanes : Anti-Cdk5 antibody [EP715Y] (ab40773) at 1/2000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CDK5 knockout HAP1 whole cell lysate
    Lane 3 : SHSY5Y whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 33 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

    Lanes 1 - 4: Merged signal (red and green). Green - ab40773 observed at 33 kDa. Red - loading control, ab130007, observed at 130 kDa.

    ab40773 was shown to specifically react with Cdk5 in wild-type HAP1 cells as signal was lost in CDK5 knockout cells. Wild-type and CDK5 knockout samples were subjected to SDS-PAGE. Ab40773 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 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.

  • ab40773 staining Cdk5 in the SH-SY5Y cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/300). a150077 was used as the secondary antibody (1/500) and nuclei stained with DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

  • Immunohistochemical analysis of frozen PFA-fixed chicken midbrain tissue cyrosection, labelling Cdk5 with ab40773 at a dilution of 1/200 incubated for 16 hours at 4°C in 0.1% PBS Triton X and 3% BSA. Blocking was with 3% BSA incubated for 1 hour at 21°C. Secondary was a goat anti-rabbit polyclonal Alexa Fluor® 488 conjugate at 1/1000.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

  • ab40773 at 1/50 immunoprecipitating Cdk5 in HeLa whole cell lysate.

    For western blotting, the primary antibody was used at a dilution of 1/1000 and a HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

  • Flow cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling Cdk5 with ab40773 (pink). Negative control Rabbit IgG (green).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

  • Paraffin-embedded human brain tissue stained for Cdk5 with ab40773 at 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Kapora E  et al. MicroRNA-505-5p functions as a tumor suppressor by targeting cyclin-dependent kinase 5 in cervical cancer. Biosci Rep 39:N/A (2019). Read more (PubMed: 31266812) »
  • Yu HP  et al. TIGAR regulates DNA damage and repair through pentosephosphate pathway and Cdk5-ATM pathway. Sci Rep 5:9853 (2015). WB . Read more (PubMed: 25928429) »
See all 7 Publications for this product

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