Anti-p21^WAF1 (58-77) (Ab-1) Mouse mAb (EA10) Fluorescein Conjugate

Purified mouse monoclonal antibody generated by immunizing F1 mice with the specified immunogen and fusing splenocytes with SP2/0 mouse myeloma cells (see application references). Recognizes the ~21 kDa p21^WAF1 protein.

Epitope: within amino acids 58-77 of human p21^WAF1

Human

full-length, recombinant, human p21^WAF1

Flow Cytometry (2 µg/ml)

Frozen Sections (5 µg/ml)

Immunoblotting (use Cat. No. OP64)

Immunofluorescence (1-5 µg/ml)

Immunoprecipitation (use Cat. No. OP64)

Paraffin Sections (5 µg/ml, see comments)

Please refer to vial label for lot-specific concentration.

In 50 mM sodium phosphate buffer, 50% glycerol.

Following initial thaw, aliquot and freeze (-20°C).

Negative Control
Unstimulated cells, unstimulated skin tissue, or SK-OV-3 cells

Positive Control
Any cell line expressing wild-type p53 (e.g. Hs27 cells or U2OS cells treated with DNA damaging agents) or UV-treated skin or colon tissue

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Maximal p21^WAF1 expression requires wild type p53 activity. Treatment of U2OS or MCF7 cells with DNA damaging agents (such as doxorubicin at 0.2 µg/ml) induces wild type p53 expression which in turn activates p21^WAF1 expression. Serum stimulation of quiescent cells will give low level p21^WAF1 expression independent of p53 expression. Untreated cells will express little p21^WAF1 and can also be used as a negative control. Cat. No. OP64F was tested in HALT cells induced by incubation at 31°C; FITC-goat anti-mouse IgG (Cat. No. DC13L) was used as a negative control. For staining paraffin sections, no pretreatment is usually needed, but some tissues may stain better after pretreating with heat or pressure cooker. In either paraffin or frozen sections of normal human colon, the non-dividing cells of colonic epithelium will stain positive for p21^WAF1 while the proliferating compartment of crypts will not stain. For immunoprecipitation and immunoblotting use Cat. No. OP64. Antibody should be titrated for optimal results in individual systems.

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Toxicity: Standard Handling (A)