Overview

  • Product name
    Anti-Cdk5 antibody [EP715Y]
    See all Cdk5 primary antibodies
  • Description
    Rabbit monoclonal [EP715Y] to Cdk5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
  • Immunogen

    Synthetic peptide within Human Cdk5 aa 250 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: HeLa whole cell lysate (ab150035). IHC-P: Human brain tissue. Flow Cyt: HeLa cells. IHC-Fr: Chicken midbrain 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 ab40773 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
IHC-Fr 1/200.
WB 1/2000 - 1/10000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/30.

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

IP 1/50.
ICC/IF 1/100 - 1/250.

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
    Observed band size: 33 kDa



    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.

  • Anti-Cdk5 antibody [EP715Y] (ab40773) at 1/10000 dilution + HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

    Predicted band size: 33 kDa
    Observed band size: 33 kDa

  • 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.

  • 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.

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

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

  • 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.

References

This product has been referenced in:
  • Yang L  et al. Thioredoxin-1 Protects Spinal Cord from Demyelination Induced by Methamphetamine through Suppressing Endoplasmic Reticulum Stress and Inflammation. Front Neurol 9:49 (2018). Read more (PubMed: 29467717) »
  • Majd S  et al. Compound C enhances tau phosphorylation at Serine396 via PI3K activation in an AMPK and rapamycin independent way in differentiated SH-SY5Y cells. Neurosci Lett 670:53-61 (2018). Read more (PubMed: 29398519) »
See all 12 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
Yes - 70% Ethanol O/N at 4C
Specification
Brain
Blocking step
Dako Block #X0909 as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 23 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Chicken Tissue (Midbrain cryosection)
Permeabilization
Yes - 0.1% PBS Triton X
Specification
Midbrain cryosection
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 12 2015

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10% SDS gel)
Sample
Human Cell lysate - whole cell (human iPS cell derived neurons (healthy/diseased))
Specification
human iPS cell derived neurons (healthy/diseased)
Treatment
quinolinic acid 72h (and non treated) 500nM
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 17 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (HeLa cell line)
Specification
HeLa cell line
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 12 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (U2OS cells)
Specification
U2OS cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 12 2013

Question
Answer

DISCOUNT CODE: *** Expiration date: April 1st, 2012 I am very pleased to hear you would like to accept our offer and test ab40773 in IHC-FoFr. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for IHC-FoFr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293)
Loading amount
20 µg
Specification
HEK 293
Gel Running Conditions
Non-reduced Denaturing (12.5)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Aug 22 2011

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