5-μm sections of paraffin embedded, formalin fixed, lung were stained for p16, p21, and p53 by immunohistochemistry as previously described. [23 (link)] Briefly, the sections were incubated in PBS containing 5% normal goat serum and 1% bovine serum albumin (BSA), then incubated with a 1:100 dilution of mouse anti-human p16 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-human p21 (Santa Cruz Biotechnology) or mouse anti-human p53 antibody (Santa Cruz Biotechnology) overnight at 4°C. The sections were washed, incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG (Santa Cruz Biotechnology). After washing, the bound peroxidase activity was detected using a diaminobenzidine (DAB) substrate kit (Vector Laboratories, Burlingame, CA).
Rabbit anti human p21
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Rabbit anti-human p21 is an antibody produced in rabbits that specifically recognizes the p21 protein from human samples. p21 is a cyclin-dependent kinase inhibitor involved in the regulation of cell cycle progression.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti human p16, by Santa Cruz Biotechnology (2 mentions)
Diaminobenzidine dab substrate kit, by Vector Laboratories (1 mentions)
Horseradish peroxidase hrp conjugated goat anti mouse or anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Mouse anti gapdh antibody, by Kangchen Biotech (1 mentions)
Mouse anti human p erk, by Santa Cruz Biotechnology (1 mentions)
Mouse anti human mmp 9, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti human bcl 2, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti human mmp 2, by Cell Signaling Technology (1 mentions)
Mouse anti human p53, by Santa Cruz Biotechnology (1 mentions)
Phototope hrp detection kit, by New England Biolabs (1 mentions)
Mouse anti human β actin, by Santa Cruz Biotechnology (1 mentions)
3 protocols using rabbit anti human p21
1
Immunohistochemical Analysis of Cell Cycle Regulators
2
Immunoprecipitation and Western Blot Analysis
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The supernatant from whole-cell lysates was harvested and immunoprecipitated using the indicated primary antibodies or non-specific IgG as a negative control, which was precleared by protein G magnetic beads at 4°C overnight. Proteins were separated on 10 or 12% SDS-PAGE gels and transferred onto nitrocellulose membranes (Bio-Rad, Hercules, CA, USA). The following antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA): mouse anti-human Gal-1, rabbit anti-human H-Ras, rabbit anti-human ERK, mouse anti-human p-ERK, rabbit anti-human c-Jun, mouse anti-human MMP-9, rabbit anti-human p21, rabbit anti-human Bcl-2, and rabbit anti-human Raf-1 antibodies. Rabbit anti-human p-AKT (Ser473), rabbit anti-human p-Raf-1 (Ser338), and rabbit anti-human MMP-2 antibodies were purchased from Cell Signaling (Danvers, MA, USA). The mouse anti-GAPDH antibody was purchased from KangChen Bio-tech (Shanghai, China). All experiments were repeated at least twice.
3
Western Blot Analysis of Cell Lysates
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Type II AECs were lysed in RIPA buffer and the cell lysates subjected to SDS-PAGE under reducing conditions and transferred to nitrocellulose membrane (Life Sciences Products, Boston, MA). [22 (link)] The membranes were washed with 50 mM Tris-HCl containing 0.5 M NaCl, 0.01% Tween 20, (TBS; pH 7.5) and incubated overnight in 5% milk containing primary antibody: mouse anti-human p16 (Santa Cruz Biotechnology), rabbit anti-human p21 (Santa Cruz Biotechnology), mouse anti-human p53 (Santa Cruz Biotechnology), or mouse anti-human β-actin (Santa Cruz Biotechnology). The membrane was washed with TBS, then incubated with secondary antibody and washed again. Immunoreactivity was detected using the phototope-HRP-detection kit (New England Biolabs, Beverly, MA).
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