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Purified Mouse Anti-Cdk4
Purified Mouse Anti-Cdk4

Immunoprecipitiation/western blot analysis of Cdk4. HeLa or 293 cell lysates were

immunoprecipitated with an antibody to Cdk4 (clone DCS-156). Proteins were separated by SDS/PAGE and then western blotted with clone DCS-156. The bands above and below the ~32 kD Cdk4 bands, represent the heavy and light chains of IgG used for immunoprecipitation.

Purified Mouse Anti-Cdk4

Western blot titration of anti-Cdk4 antibody. HeLa or 293 cell lysates were probed with 6 ug/ml (lanes 1 and 4), 2.0 µg/ml (lanes 2 and 4) or 0.5 µg/ml (lanes 3 and 5) of antibody (clone DCS-156). The antibody identifies Cdk4 at ~32 kD.

Immunoprecipitiation/western blot analysis of Cdk4. HeLa or 293 cell lysates were

immunoprecipitated with an antibody to Cdk4 (clone DCS-156). Proteins were separated by SDS/PAGE and then western blotted with clone DCS-156. The bands above and below the ~32 kD Cdk4 bands, represent the heavy and light chains of IgG used for immunoprecipitation.

Western blot titration of anti-Cdk4 antibody. HeLa or 293 cell lysates were probed with 6 ug/ml (lanes 1 and 4), 2.0 µg/ml (lanes 2 and 4) or 0.5 µg/ml (lanes 3 and 5) of antibody (clone DCS-156). The antibody identifies Cdk4 at ~32 kD.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Recombinant Human Cdk4
Western blot (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required), Immunoprecipitation (Tested During Development)
32 kDa
0.5 mg/ml
AB_397299
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Applications include western blot analysis (0.5-2.0 µg/ml) and immunoprecipitation (1.0 µg/one million cells). The antibody has also been used for immunohistochemistry of formalin-fixed, paraffin-embedded tissue sections, but this application is not routinely tested at BD Biosciences Pharmingen. HeLa (ATCC CCL 2), 293 (ATCC CRL 1573), or NIH/3T3 (ATCC CRL 1658) cells are suggested as positive controls.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
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Antibody Details
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DCS-156

Cyclins, cyclin-dependent kinases (Cdks), and cyclin-dependent kinase inhibitors (CdkIs) are essential for cell-cycle control in eukarytotes. Cyclins, regulatory subunits, bind to cyclin-dependent kinases (Cdks), catalytic subunits, to form active cyclin-Cdk complexes. Cdk subunits by themselves are inactive and binding to a cyclin is required for their activity. Cyclins A, B1, D and E undergo periodic synthesis and degradation, thereby providing a mechanism to regulate Cdk activity throughout the cell cycle. Additionally, Cdk activity is further regulated by activating and inhibitory phosphorylations, and small proteins (p15, p16, p18, p19, p21 and p27), called inhibitors of Cdk activity, that bind to cyclins, Cdks, or cyclin- Cdk complexes. Cdk4 was originally called PSK-J3, and following demonstration of its association with D-type cyclins, became known as Cdk4. D-type cyclins also associate with Cdks 2 and 5, although Cdk4 appears to be the most abundant partner. The D-type cyclins (D1, D2, and D3) are expressed in response to growth factors or mitogens, and rapidly degrade when mitogens are withdrawn. D cyclins appear to promote G0 to G1 transitions and the rate of G1 progression. For example, cyclin D-Cdk4 and cyclin D-Cdk6 complexes phosphorylate the retinoblastoma protein (Rb) which removes the G1 phase block caused by underphosphorylated Rb. Cdk4 has a molecular weight of ~32 kD.  Clone DCS-156 recognizes human and mouse Cdk4.  A recombinant protein fragment from the C-terminal end of human Cdk4 was used as immunogen.

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Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
559677 Rev.3
Citations & References
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Development References (2)

  1. Johnson DG, Walker CL. Cyclins and cell cycle checkpoints. Annu Rev Pharmacol Toxicol. 1999; 39:295-312. (Biology). View Reference
  2. Matsushime H, Ewen ME, Strom DK, et al. Identification and properties of an atypical catalytic subunit (p34PSK-J3/cdk4) for mammalian D type G1 cyclins. Cell. 1992; 71(2):323-334. (Biology). View Reference
559677 Rev. 3

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.