Rabbit CDK4 Polyclonal Antibody | anti-CDK4 antibody

Phospho-CDK4 (Thr172) Antibody

Cat. Num. AAA9614008

• Gene Names: CDK4; CMM3; PSK-J3
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Zebrafish (85%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (100%), Xenopus (92%)
• Applications: Western Blot, Immunohistochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: CDK4; Polyclonal Antibody; Phospho-CDK4 (Thr172) Antibody; Cdk 4; cdk4; CDK4 protein; CDK4_HUMAN; Cell division kinase 4; Cell division protein kinase 4; CMM 3; CMM3; Crk3; Cyclin dependent kinase 4; Cyclin-dependent kinase 4; Melanoma cutaneous malignant 3; MGC14458; p34 cdk4; PSK J3; PSK-J3; anti-CDK4 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Zebrafish (85%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (100%), Xenopus (92%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-CDK4 (Thr172) Antibody detects endogenous levels of CDK4 only when phosphorylated at Thr172.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-CDK4 antibody: Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IHC: 1:50-1:200; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human CDK4 around the phosphorylation site of Thr172.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: Phosphorylation 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.
• Subunit Structure: Component of the D-CDK4 complex, composed of CDK4 and some D-type G1 cyclin (CCND1, CCND2 or CCND3). Interacts directly in the complex with CCND1, CCND2 or CCND3. Interacts with SEI1 and ZNF655. Forms a ternary complex, cyclin D-CDK4-CDKN1B, involved in modulating CDK4 enzymatic activity. Interacts directly with CDKN1B (phosphorylated on 'Tyr-88' and 'Tyr-89'); the interaction allows assembly of the cyclin D-CDK4 complex, Thr-172 phosphorylation, nuclear translocation and enhances the cyclin D-CDK4 complex activity. CDK4 activity is either inhibited or enhanced depending on stoichiometry of complex. The non-tyrosine-phosphorylated form of CDKN1B prevents T-loop phosphorylation of CDK4 producing inactive CDK4. Interacts (unphosphorylated form) with CDK2. Also forms ternary complexes with CDKN1A or CDKN2A. Interacts directly with CDKN1A (via its N-terminal); the interaction promotes the assembly of the cyclin D-CDK4 complex, its nuclear translocation and promotes the cyclin D-dependent enzyme activity of CDK4. Interacts with CCND1; the interaction is prevented with the binding of CCND1 to INSM1 during cell cycle progression. Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones CDC37, PPP5C, TSC1 and client protein TSC2, CDK4, AKT, RAF1 and NR3C1; this complex does not contain co-chaperones STIP1/HOP and PTGES3/p23. Interacts with CEBPA (when phosphorylated). Interacts with FNIP1 and FNIP2.
• Similarity: Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
• Subcellular Location: Cytoplasm. Nucleus. Nucleus membrane.; Note: 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.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of CDK4 (Phospho-Thr172) using serum treated K562 whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunohistochemistry (IHC)

(Staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human esophageal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human normal tissues adjacent to esophageal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/200 staining Rat intestinal tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/200 staining Mouse liver tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/200 staining Human prostate cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Related Product Information for anti-CDK4 antibody

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.

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: 34kDa
• NCBI Official Full Name: cyclin-dependent kinase 4
• NCBI Official Synonym Full Names: cyclin-dependent kinase 4
• NCBI Official Symbol: CDK4
• NCBI Official Synonym Symbols: CMM3; PSK-J3
• NCBI Protein Information: cyclin-dependent kinase 4; cell division protein kinase 4
• UniProt Protein Name: Cyclin-dependent kinase 4
• Protein Family: Cyclin-dependent kinase
• UniProt Gene Name: CDK4
• UniProt Entry Name: CDK4_HUMAN