CDK4 antibody (N-Term)

Catalog No. ABIN967650

This anti-CDK4 antibody is a Mouse Monoclonal antibody detecting CDK4 in WB, IP, IHC (fro) and BI. Suitable for Human, Mouse, Rat and Dog. This Primary Antibody has been cited in 2+ publications.

Quick Overview for CDK4 antibody (N-Term) (ABIN967650)

Target: CDK4 (Cyclin-Dependent Kinase 4 (CDK4))

Reactivity: Human, Mouse, Rat, Dog

Host: Mouse

Clonality: Monoclonal

Conjugate: This CDK4 antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Frozen Sections) (IHC (fro)), BioImaging (BI)

Clone: DCS-35

Product Details anti-CDK4 Antibody

Binding Specificity: N-Term

Brand: BD Pharmingen™

Cross-Reactivity: Dog (Canine), Rat (Rattus), Mouse (Murine)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. 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.
5. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Cdk4

Isotype: IgG1 kappa

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C.

References for anti-CDK4 antibody (ABIN967650)

Johnson, Walker: "Cyclins and cell cycle checkpoints." in: Annual review of pharmacology and toxicology, Vol. 39, pp. 295-312, (1999) (PubMed).

Matsushime, Ewen, Strom, Kato, Hanks, Roussel, Sherr: "Identification and properties of an atypical catalytic subunit (p34PSK-J3/cdk4) for mammalian D type G1 cyclins." in: Cell, Vol. 71, Issue 2, pp. 323-34, (1992) (PubMed).

Target Details for CDK4

Target: CDK4 (Cyclin-Dependent Kinase 4 (CDK4))

Alternative Name: Cdk4

Background: Cyclins, cyclin-dependent kinases (Cdks), and cyclin-dependent kinase inhibitors (CdkIs) are essential for cell-cycle control in eukaryotes (reviewed in 1). 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 CdkIs, that bind to cyclins, Cdks, or cyclin-Cdk complexes. Cdk4 was originally called PSK-J3,2 and following demonstration of its association with D-type cyclins, became known as Cdk4.2 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 ~33 kD.

Molecular Weight: 33 kDa

Pathways: Cell Division Cycle, Mitotic G1-G1/S Phases, Regulation of Cell Size