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Immunofluorescent analysis of p21 Waf1/Cip1 (green) in HeLa cells either left untreated (left panel) or treated with 5uM Camptothecin for 3 hours (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Dog, Human, Primate|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues near the carboxy-terminus of human p21|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Contains||<0.02% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
It is not recommended to aliquot this antibody.
This antibody is not cross-reactive with other CDK inhibitors.
This gene encodes a potent cyclin-dependent kinase inhibitor. The encoded protein binds to and inhibits the activity of cyclin-CDK2 or -CDK4 complexes, and thus functions as a regulator of cell cycle progression at G1. The expression of this gene is tightly controlled by the tumor suppressor protein p53, through which this protein mediates the p53-dependent cell cycle G1 phase arrest in response to a variety of stress stimuli. This protein can interact with proliferating cell nuclear antigen (PCNA), a DNA polymerase accessory factor, and plays a regulatory role in S phase DNA replication and DNA damage repair. This protein was reported to be specifically cleaved by CASP3-like caspases, which thus leads to a dramatic activation of CDK2, and may be instrumental in the execution of apoptosis following caspase activation. Two alternatively spliced variants, which encode an identical protein, have been reported.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Role of stress-activated OCT4A in the cell fate decisions of embryonal carcinoma cells treated with etoposide.
MA5-14949 was used in western blot to elucidate the mechanism of p53-dependent upregulation of OCT4A and p21Cip1.
|Huna A,Salmina K,Erenpreisa J,Vazquez-Martin A,Krigerts J,Inashkina I,Gerashchenko BI,Townsend PA,Cragg MS,Jackson TR||Cell cycle (Georgetown, Tex.) (14:2969)||2015|
CDK-interacting protein 1; CDK-interaction protein 1; cyclin-dependent kinase inhibitor 1; cyclin-dependent kinase inhibitor 1A (p21, Cip1); DNA synthesis inhibitor; melanoma differentiation associated protein 6; melanoma differentiation-associated protein 6; OTTHUMP00000016298; p21Cip1/Waf1; wild-type p53-activated fragment 1
CAP20; CDKN1; CDKN1A; CIP1; MDA-6; MDA6; P21; p21CIP1; PIC1; SDI1; WAF1