Invitrogen

CDK4 Recombinant Rabbit Monoclonal Antibody (SD20-42)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

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Datasheet

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Datasheet

Protocols

Questions & Answers

Tech Support

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CDK4 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry/Immunofluorescence analysis of CDK4 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with CDK4 Recombinant Monoclonal Antibody (Product # MA5-41178) at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1:1,000 dilution. The nuclear counter stain is DAPI (blue).

CDK4 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of CDK4 in paraffin-embedded human stomach carcinoma tissue. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with CDK4 Recombinant Monoclonal Antibody (Product # MA5-41178) at a dilution of 1:50 for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

CDK4 Antibody in Western Blot (WB) — Knockout of CDK4 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1007917_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CDK4 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) andHeLa CDK4 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-CDK4 Recombinant Rabbit Monoclonal Antibody (SD20-42) (Product # MA5-41178, 1:5000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CDK4.

CDK4 Antibody in Flow Cytometry (Flow) — Flow Cytometry analysis of CDK4 in MCF-7 cells. The cells were fixed, permeabilized and stained with CDK4 Recombinant Monoclonal Antibody (Product # MA5-41178) at a dilution of 1:50 (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1:1,000 dilution for 30 minutes. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

CDK4 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inCDK4 KO cell line compared to control cell line using Anti-CDK4 Recombinant Rabbit Monoclonal Antibody (SD20-42) (Product # MA5-41178).

Product Details

MA5-41178

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000-1:5,000

Immunohistochemistry (Paraffin) (IHC (P))

1:50-1:200

Immunocytochemistry (ICC/IF)

1:100-1:500

Flow Cytometry (Flow)

1:50-1:100

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Expression System

HEK293 cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

SD20-42

Immunogen

Synthetic peptide within C-terminal human Cdk4

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 0.05% BSA, 40% glycerol

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2898932

Target Information

CDK4 is a member of the Ser/Thr protein kinase family. This protein is highly similar to the gene products of S. cerevisiae cdc28 and S. pombe cdc2. It is a catalytic subunit of the protein kinase complex that is important for cell cycle G1 phase progression. The activity of this kinase is restricted to the G1-S phase, which is controlled by the regulatory subunits D-type cyclins and CDK inhibitor p16(INK4a). This kinase was shown to be responsible for the phosphorylation of retinoblastoma gene product (Rb). Mutations in this gene as well as in its related proteins including D-type cyclins, p16(INK4a) and Rb were all found to be associated with tumorigenesis of a variety of cancers. Multiple polyadenylation sites of the gene have been reported.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Cell division protein kinase 4; Cyclin-dependent kinase 4; MGC14458; PSK-J3

Gene Aliases: CDK4; CMM3; PSK-J3

UniProt ID: (Human) P11802

Entrez Gene ID: (Human) 1019

Molecular Function: non-receptor serine/threonine protein kinase

Function(s)

G1/S transition of mitotic cell cycle lens development in camera-type eye protein phosphorylation signal transduction circadian rhythm positive regulation of cell proliferation response to toxic substance response to lead ion regulation of gene expression positive regulation of G2/M transition of mitotic cell cycle organ regeneration response to testosterone response to drug positive regulation of apoptotic process positive regulation of translation positive regulation of cell cycle positive regulation of cell size regulation of protein kinase activity positive regulation of fibroblast proliferation cell division response to hyperoxia negative regulation of cell cycle arrest

Process(es)

cyclin-dependent protein serine/threonine kinase activity protein binding ATP binding cyclin-dependent protein serine/threonine kinase regulator activity cyclin binding protein complex binding