Rabbit anti-Human CDK4 Monoclonal Antibody | anti-CDK4 antibody

[KO Validated] CDK4 Rabbit PolymAb

Cat. Num. AAA28563

• Reactivity: Human
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, Immunoprecipitation, ELISA
• Purity: Affinity purification
• Synonyms: CDK4, Antibody; [KO Validated] CDK4 Rabbit PolymAb; CMM3; PSK-J3; anti-CDK4 antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Monoclonal
• Isotype: IgG
• Purity/Purification: Affinity purification
• Form/Format: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
• Sequence: FAEMFRRKPLFCGNSEADQLGKIFDLIGLPPEDDWPRDVSLPRGAFPPRGPRPVQSVVPEMEESGAQLLLEMLTFNPHKRISAFRALQHSYLHKDEGNPE

• Applicable Applications for anti-CDK4 antibody: Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunofluorescence (IF), Immunocytochemistry (ICC), Immunoprecipitation (IP), ELISA (EIA)
• Application Notes: WB: 1:1000-1:12000; IHC-P: 1:200-1:800; IF/ICC: 1:200-1:800; IP: 0.5ug-4ug antibody for 200ug-400ug extracts of whole cells; ELISA: Recommended starting concentration is 1ug/mL.; Please optimize the concentration based on your specific assay requirements.
• Cross Reactivity: Human, Mouse, Rat
• Immunogen: A synthetic peptide corresponding to a sequence within amino acids 204-303 of human CDK4 (NP_000066.1).
• Preparation and Storage: Store at -20 degree C. Avoid freeze/thaw cycles.

IP (Immunoprecipitation)

(Immunoprecipitation of CDK4 from 300 µg extracts of HeLa cells was performed using 2 µg of [KO Validated] CDK4 Rabbit PolymAb® (AAA28563). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at a dilution of 1:3000.)

ICC (Immunocytochemistry)

(Confocal imaging of paraffin-embedded Rat lung tissue using [KO Validated] CDK4 Rabbit PolymAb® (A23521-PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.)

ICC (Immunocytochemistry)

(Confocal imaging of HeLa cells using [KO Validated] CDK4 Rabbit PolymAb® (A23521-PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.)

ICC (Immunocytochemistry)

(Confocal imaging of HeLa cells using [KO Validated] CDK4 Rabbit PolymAb® (A23521-PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with ?-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.)

IHC (Immunohistochemistry)

(Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.)

IHC (Immunohistochemistry)

(Immunohistochemistry analysis of paraffin-embedded Human breast cancer tissue using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.)

IHC (Immunohistochemistry)

(Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.)

WB (Western Blot)

(Western blot analysis of various lysates using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at 1:1000 dilution incubated overnight at 4?.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25 ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Exposure time: 30s.)

WB (Western Blot)

(Western blot analysis of lysates from wild type (WT) and CDK4 knockout (KO) HeLa cells using [KO Validated] CDK4 Rabbit PolymAb® (AAA28563) at 1:3000 dilution incubated overnight at 4?.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25 ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Exposure time: 30s.)

Related Product Information for anti-CDK4 antibody

The protein encoded by this gene 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 this gene have been reported.

Product Categories/Family for anti-CDK4 antibody

Epigenetics Nuclear Signaling; Protein phosphorylation; Cancer; Signal Transduction; Kinase; Serine threonine kinases; Cell Biology Developmental Biology; Cell Cycle; Cell Cycle Control-G1 S Checkpoint; Antibodies

NCBI and Uniprot Product Information

• NCBI GI #: 4502735
• NCBI GeneID: 1019
• NCBI Accession #: NP_000066.1
• NCBI GenBank Nucleotide #: NM_000075.3
• UniProt Accession #: P11802; O00576; Q6FG61
• Molecular Weight: Calculated MW: 34kDa; Observed MW: 34kDa
• UniProt Protein Name: Cyclin-dependent kinase 4
• UniProt Gene Name: CDK4
• UniProt Entry Name: CDK4_HUMAN