This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide at 0.5 mg/ml.
Preparation:
The antibody was purified by affinity chromatography.
Concentration:
0.5 mg/ml
Storage & Handling:
Upon receipt, store undiluted at 4° C.
Application:
WB - Quality Tested IHC(Paraffin)1*, ELISA * - *This application has been reported in the literature.
Recommended Usage:
Each lot of this antibody is quality control tested by Western blotting. For Western blotting, suggested working dilution(s): Use 5 μg per 5 ml antibody dilution buffer for each mini-gel. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: immunohistochemistry1 of formalin-fixed, paraffin-embeded tissue sections, and ELISA.
Application References:
1. Kolar, Z., et al., 2000. Mol. Pathol. 53:15.
MCF7 cell lysates were resolved by electrophoresis, transferred to nitrocellulose, and probed with monoclonal antibody against p21. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.
Description:
p21, also known as cyclin-dependent kinase inhibitor 1A (CDKN1A), WAF-1/CIP-1, and MDA 6, is a protein containing a nuclear localization sequence that inhibits cyclin-kinase activity. This nuclear protein can also be found in the cytoplasm under certain conditions such as overexpression of the BRAP protein. p21 inhibits cyclin-kinase activity, serving as the effector of p53 cell cycle control. Over-expression of p21 can rescue cells from apoptosis and is thought to be required to maintain hematopoietic stem cell quiescence. p21 is tightly controlled at the transcriptional level by p53. The p21 protein is phosphorylated by Akt1, PIM1, and JNK1 and interacts with a variety of proteins involved in cell-cycle control, and DNA-damage responses including casein kinase II beta, cyclin D1, cyclin E1, proliferating cell nuclear antigen, and GADD45. The WA-1 antibody recognizes human and monkey p21 and has been shown to be useful for Western blotting immunohistochemistry using formalin-fixed, paraffin-embedded tissues, and ELISA.
Other Names:
CDKN1A, cyclin-dependent kinase inhibitor 1A, Wild-type p53-activated fragment 1, (WAF-1), cdk interacting protein (CIP1), melanoma differentiation associated protein 6 (MDA 6)
Structure:
Contains nuclear localization sequence, CDKN1A is proteolytically cleaved by caspase 3 at residue 112 and 149
Distribution:
Nuclear protein; cytoplasmic when BRAP overexpressed
Function:
Inhibitor of cyclin-kinase activity, probably serves as the effector of p53 cell cycle control. Over-expression of p21 can rescue cells from apoptosis and is thought to be required to maintain hematopoietic stem cell quiescence.
Regulation:
Tightly regulated at the transcriptional level by p53
Modification:
Phosphorylation (Thr57, Ser130, Thr145)
Interaction:
Casein kinase II beta, cyclin D1, cyclin E1, proliferating cell nuclear antigen, GADD45, and many others
Antigen References:
1. Bunz, F., et al., 1998. Science 282:1497. 2. Cheng, T., et al., 2000. Science 287:1804. 3. El-Diery, W. S., et al., 1993. Cell 75:817. 4. Harper, J. W., et al., 1993. Cell 75:805.
