Anti-Cdk4 antibody (ab95255)
Key features and details
- Rabbit polyclonal to Cdk4
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Cdk4 antibody
See all Cdk4 primary antibodies -
Description
Rabbit polyclonal to Cdk4 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- This antibody gave a positive signal in both MCF7 and HeLa whole cell lysates.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab95255 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species WB HumanAll applications PigApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).Target
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Function
Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex. -
Involvement in disease
Defects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B. -
Cellular localization
Cytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 1019 Human
- Entrez Gene: 100144492 Pig
- Omim: 123829 Human
- SwissProt: P11802 Human
- SwissProt: P79432 Pig
- Unigene: 95577 Human
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Alternative names
- Cdk 4 antibody
- cdk4 antibody
- CDK4 protein antibody
see all
Images
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Western blot - Anti-Cdk4 antibody (ab95255)
Lanes 1: Wild-type HAP1 cell lysate (20 µg)
Lanes 2: CDK4 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Merged (red and green) signal. Green signal from target – ab95255 observed at 34 kDa. Red signal from loading control – ab8226 observed at 42 kDaab95255 was shown to react with CDK4 when CDK4 knockout samples were used, along with additional cross-reactive bands. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab95255 and ab8226 (loading control to beta actin) were diluted 1 µg/mL and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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Western blot - Anti-Cdk4 antibody (ab95255)Anti-Cdk4 antibody (ab95255) at 1 µg/ml + MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
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: 33 kDa
Observed band size: 33 kDa
Additional bands at: 110 kDa, 16 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
A positive signal was also observed within HeLa whole cell lysate. Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab95255 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-Cdk4 antibody (ab95255)
Lanes 1: Wild-type HAP1 cell lysate (20 µg)
Lanes 2: CDK4 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Merged (red and green) signal. Green signal from target – ab95255 observed at 34 kDa. Red signal from loading control – ab8226 observed at 42 kDaab95255 was shown to react with CDK4 when CDK4 knockout samples were used, along with additional cross-reactive bands. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab95255 and ab8226 (loading control to beta actin) were diluted 1 µg/mL and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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Western blot - Anti-Cdk4 antibody (ab95255)Anti-Cdk4 antibody (ab95255) at 1 µg/ml + MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
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: 33 kDa
Observed band size: 33 kDa
Additional bands at: 110 kDa, 16 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
A positive signal was also observed within HeLa whole cell lysate. Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
