CDK4 Antibody [Unconjugated]

Western blot shows lysates of A431 human epithelial carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for CDK4 at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CDK4 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

CDK4 was detected in immersion fixed RAW264.7 cells using Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cells, and MCF‑7 human breast cancer cells, loaded at 0.2 mg/mL. A specific band was detected for CDK4 at approximately 38 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CDK4 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human CDK4
  Met1-Glu303
  Accession # P11802
• Specificity: Detects human and mouse CDK4 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CDK4
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CDK4 Antibody [Unconjugated]

• CDK4
• Cell division protein kinase 4
• CMM3
• cyclin-dependent kinase 4
• EC 2.7.11
• EC 2.7.11.22
• melanoma cutaneous malignant, 3
• MGC14458
• PSK-J3
• PSK-J3cell division kinase 4

Background

CDK4 (cyclin-dependent kinase 4) is a 34 kDa member of the CMGC Ser/Thr protein kinase family of enzymes. Within the cell, pRb/retinoblastoma protein is normally unphosphorylated and suppresses cell growth at the G1 stage of the cell cycle. When mitosis is required, CDK4 is phosphorylated, forms a complex with cyclin D1, 2 or 3, and phosphorylates pRb/retinoblastoma protein. This inactivates pRb and cells initiate DNA synthesis. Human CDK4 is 303 amino acids (aa) in length. It contains one protein kinase domain (aa 6‑295) and an activating phosphorylation site at Thr172. There are three potential variants; one that shows an alternate start site at Met75, a second that shows a two aa substitution for aa 89‑211, and a third that shows a 23 aa substitution for aa 89‑303. Full-length human and mouse CDK4 are 95% aa identical.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CDK4 Antibody (AF5254)(2)

Publications using AF5254

• L Zhang, Y Li, C Hu, Y Chen, Z Chen, ZS Chen, JY Zhang, S Fang CDK6-PI3K signaling axis is an efficient target for attenuating ABCB1/P-gp mediated multi-drug resistance (MDR) in cancer cells Molecular Cancer, 2022-04-22;21(1):103. 2022-04-22 [PMID: 35459184] (Western Blot, Human)
• JM Oh, J Lee, WT Im, S Chun Ginsenoside Rk1 Induces Apoptosis in Neuroblastoma Cells Through Loss of Mitochondrial Membrane Potential and Activation of Caspases Int J Mol Sci, 2019-03-11;20(5):. 2019-03-11 [PMID: 30862004] (Western Blot, Human)

Bioinformatics

• Gene Symbol: CDK4
• Uniprot:
  • P11802()