CDKN1B antibody

Catalog No. ABIN967391

Product Details anti-CDKN1B Antibody

Target: CDKN1B (Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B))

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This CDKN1B antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP), BioImaging (BI)

Brand: BD Pharmingen™

Cross-Reactivity: Human

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us 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.
4. 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.
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: Mouse p27 [Kip1] (full-length) Recombinant Protein

Clone: G173-524

Isotype: IgG1

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-CDKN1B antibody (ABIN967391)

Gorospe, Liu, Xu, Chrest, Holbrook: "Inhibition of G1 cyclin-dependent kinase activity during growth arrest of human breast carcinoma cells by prostaglandin A2." in: Molecular and cellular biology, Vol. 16, Issue 3, pp. 762-70, (1996) (PubMed).

Nourse, Firpo, Flanagan, Coats, Polyak, Lee, Massague, Crabtree, Roberts: "Interleukin-2-mediated elimination of the p27Kip1 cyclin-dependent kinase inhibitor prevented by rapamycin." in: Nature, Vol. 372, Issue 6506, pp. 570-3, (1995) (PubMed).

Polyak, Kato, Solomon, Sherr, Massague, Roberts, Koff: "p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest." in: Genes & development, Vol. 8, Issue 1, pp. 9-22, (1994) (PubMed).

Toyoshima, Hunter: "p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21." in: Cell, Vol. 78, Issue 1, pp. 67-74, (1994) (PubMed).

Sherr: "Mammalian G1 cyclins." in: Cell, Vol. 73, Issue 6, pp. 1059-65, (1993) (PubMed).

Target Details for CDKN1B

Target: CDKN1B (Cyclin-Dependent Kinase Inhibitor 1B (p27, Kip1) (CDKN1B))

Alternative Name: p27 Kip1 (CDKN1B Products)

Synonyms: CDKN4 antibody, KIP1 antibody, MEN1B antibody, MEN4 antibody, P27KIP1 antibody, cdkn1b antibody, sb:cb611 antibody, wu:fb63g10 antibody, wu:fb64g10 antibody, wu:fe18e03 antibody, CDKN1B antibody, Kip1 antibody, p27 antibody, FAM14D antibody, ISG12 antibody, ISG12A antibody, P27 antibody, Cdki1b antibody, AA408329 antibody, AI843786 antibody, p27Kip1 antibody, cdkn1bl antibody, cyclin dependent kinase inhibitor 1B antibody, cyclin-dependent kinase inhibitor 1Bb antibody, interferon alpha inducible protein 27 antibody, cyclin-dependent kinase inhibitor 1B antibody, cyclin-dependent kinase inhibitor 1Ba antibody, CDKN1B antibody, cdkn1bb antibody, IFI27 antibody, Cdkn1b antibody, cdkn1ba antibody

Background: Cyclins and cyclin-dependent kinases (cdks) are evolutionarily conserved proteins that are essential for cell-cycle control in eukaryotes. Cyclins (regulatory subunits) bind to cdks (catalytic subunits) to form complexes that regulate the progression of the cell cycle. These complexes are regulated by activating and inhibitory phosphorylation events, as well as by interactions with small proteins that bind to cyclins, cdks, or cyclin-cdk complexes. These include p15, p16, p18, p19, p21 and p27 [Kip1]. p27 [Kip1] has been shown to inhibit the activity of multiple cyclin-cdk complexes, including cyclin D-cdk4, cyclin E-cdk2 and cyclin A-cdk2. p27 [Kip1] is a 27 kD protein which shares N-terminal sequence homology with p21, and like p21, p27 [Kip1] contains a nuclear localization signal in its C-terminal region. IL-2 activation of T cells has been reported to lead to a decrease in p27 [Kip1] and entry into S phase. Removal of IL-2 from T cell cultures results in increased levels of p27 [Kip1] and cell quiescence.

Molecular Weight: 27 kDa

Pathways: Cell Division Cycle, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Positive Regulation of Peptide Hormone Secretion, Negative Regulation of Hormone Secretion, Sensory Perception of Sound, Mitotic G1-G1/S Phases, DNA Replication, Positive Regulation of Endopeptidase Activity, Synthesis of DNA, Autophagy