SHK (purity > 98%) was purchased from Chengdu Must Bio-technology Co., Ltd. (Chengdu, China). MTT and NAC were purchased from Sigma-Aldrich (St. Louis, MO, United States). PI staining was purchased from Biosciences (BD Biosciences, Franklin Lakes, NJ, United States). β-actin, MEK-1, p53 and p21waf primary antibodies and HRP-conjugated secondary antibody were purchased from Santa Cruz Biotechnologies Inc. (Santa Cruz, CA, United States). Bcl-2, Bax, Rb, Cleaved Caspase-3, Ras, H3K36me2, p-p53, p-H2A.X, Histone H3 and Cyclin D1 antibodies were purchased from Cell Signaling Technology Inc. (Beverly, MA, United States). Kdm2b antibody was purchased from Millipore (Burlington, MA, United States). p16 antibody was purchased from Abcam (Cambridge, United Kingdom). Specific small interfering RNA (siRNA) of p21 and riboFECTTM CP Transfection Kit (166T) were purchased from Ribobio, Co., Ltd (Guangdong, China). Two human LAC cells (A549 and H1299) were purchased from ATCC (Manassas, VA, United States). A549 and H1299 cells were maintained in RPMI-1640 medium with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin solution, and cultured in a humidified atmosphere at 37°C with 5% CO2.
P16 antibody
Manufactured by Abcam
Sourced in United Kingdom, United States
The P16 antibody is a laboratory research tool used to detect the presence and expression levels of the p16 protein. The p16 protein is a tumor suppressor involved in cell cycle regulation. This antibody can be used in various immunoassay techniques, such as Western blotting and immunohistochemistry, to analyze p16 expression in cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
P h2ax, by Cell Signaling Technology (1 mentions)
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
H3k36me2, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
P27kip1, by Abcam (1 mentions)
P21 antibody, by Abcam (1 mentions)
Nrf2 antibody, by Cell Signaling Technology (1 mentions)
Ki67 antibody, by Abways (1 mentions)
Pcna antibody, by Proteintech (1 mentions)
Keap1 antibody, by Proteintech (1 mentions)
Trim25 antibody, by Proteintech (1 mentions)
Sftpc antibody, by ABclonal (1 mentions)
4 protocols using p16 antibody
1
Lung Cancer Cell Line Characterization
2
Immunohistochemical Analysis of Lacrimal Sac
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All lacrimal sac tissues were acquired from DCR and fixed with 4% paraformaldehyde for 24 hours. After being dehydrated, the specimens were embedded in paraffin. Then, the 4 µm serial sections were cut and blocked with 3% BSA, followed by incubating with P16 Antibody (Abcam, Cat #ab108349), P27 KIP1 (Abcam, Cat #ab32034) overnight at 4°C. After that, the sections were washed by PBS 3 times and incubated with secondary antibody for 1 hour at room temperature. Diaminobenzidine was then used to detect the immunoreactivity and the sections were washed with flowing water. Finally, after hematoxylin re-staining and washing, the sections were observed under a microscope.
3
Immunofluorescence Staining of Cellular Proteins
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Tissue sections were deparaffinized and endogenous peroxidase was blocked with 3% H2O2. Sections were incubated in 5% albumin bovine V (BSA, Solarbio) for 1 h. The cells were washed three times with PBS and fixed with 4% paraformaldehyde for 15 min. After three washes with PBS, the cells were permeated with 0.1% Triton X-100 for 15 min. After three washes with PBS, the cells were incubated with 5% bovine serum albumin for 30 min at room temperature and then incubated with p21 antibody (1:300, Abcam, Cambridge, MA, USA), p16 antibody (1:200, Abcam), SFTPC antibody (1:200, Abclonal, Wuhan, China), Keap1 antibody (1:300, Proteintech, Wuhan, China), Nrf2 antibody (1:300, Cell Signaling Technology, USA), PCNA antibody (1:200, Proteintech), Ki67 antibody (1:200, Abways, Shanghai, China), or Trim25 antibody (1:200, Proteintech) at 4 °C overnight. The next day, after washing with PBS, fluorescein-labeled secondary antibodies (1:300, Abcam) for immunofluorescence were incubated. Nuclei were counterstained with DAPI (Invitrogen, USA). The samples were washed three times with PBS and photographed with fluorescence microscopy (Nikon, Japan).
4
Quantitative PCR and Western Blot Analysis of Islet Proteins
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Quantitative PCR (qPCR) from cDNA made from DNase-digested RNA was performed with genespecific primers with SYBR green mix and normalized to the expression of housekeeping genes (TopI, Actb, and Ppia). The sequences of the primers used are available in extended information table.
Islets were isolated from each mouse individually as described above and 150-200 islets were snap frozen immediately. Western blotting was performed, as described before 31 . Briefly, total protein was extracted from 150-200 islets mice using a protein lysis buffer with a cocktail of protease inhibitors and separated by a standard SDS-PAGE and transferred to a nitrocellulose membrane. Tead1 antibody (rabbit polyclonal, 1:10,000; Abcam), p16 antibody (mouse monoclonal, 1:1,000; Abcam), Pdx1 (rabbit polyclonal, 1:10,000; BCBC), and β-actin antibody (rabbit monoclonal-horseradish peroxidase conjugate, 1:10,000; Cell Signaling), as a housekeeping control, were visualized by enhanced chemiluminescence (Pierce).
Islets were isolated from each mouse individually as described above and 150-200 islets were snap frozen immediately. Western blotting was performed, as described before 31 . Briefly, total protein was extracted from 150-200 islets mice using a protein lysis buffer with a cocktail of protease inhibitors and separated by a standard SDS-PAGE and transferred to a nitrocellulose membrane. Tead1 antibody (rabbit polyclonal, 1:10,000; Abcam), p16 antibody (mouse monoclonal, 1:1,000; Abcam), Pdx1 (rabbit polyclonal, 1:10,000; BCBC), and β-actin antibody (rabbit monoclonal-horseradish peroxidase conjugate, 1:10,000; Cell Signaling), as a housekeeping control, were visualized by enhanced chemiluminescence (Pierce).
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