Staining was performed with the VECTASTAIN ABC Elite kit (Vector Labs) according to the accompanying protocol. Antibodies for α-SMA (19245), EZH2 (5246), RAS (67648), ZMIZ1 (89500), and ARID1B (65747) were purchased from Cell Signaling Technology. Ki67 antibody (ab 16667) was purchased from Abcam. Briefly, slides were deparaffinized in three changes of xylene, 15 min each, and rehydrated in changes of 100% ethanol, 95% ethanol, and 75% ethanol for 5 min each. Sections were then washed in water twice, 5 min per wash. Antigens were unmasked by boiling the slides for 15–20 min in antigen unmasking citrate buffer (Cell Signaling Technology, 14746) and cooled down to room temperature. Endogenous peroxidase activity was quenched by incubating the slides in 3% hydrogen peroxide/H2O. Slides were washed in H2O and PBS and subsequently blocked with normal blocking serum for 30 min at room temperature. Primary antibody incubation was performed overnight at 4 °C. Following three washes in PBS, tissues were incubated in secondary biotinylated antibody for 1 h at room temperature. Tissues were treated with the avidin/biotin complex reaction according to the manufacturer’s protocol. Slides were finally washed with PBS, changed into water, and developed using the ImmPACT DAB HRP substrate kit (Vector Labs SK-4105).
Ki67 antibody ab 16667
Manufactured by Abcam
The Ki67 antibody (ab 16667) is a laboratory reagent used for the detection of the Ki67 protein, a cellular marker for proliferation. This antibody can be used in various immunoassay techniques, such as immunohistochemistry and flow cytometry, to identify and quantify proliferating cells in biological samples.
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Lab products found in correlation
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Immpact dab hrp substrate kit, by Vector Laboratories (1 mentions)
Pelp1 antibody a300 180a, by Fortis Life Sciences (1 mentions)
Fulvestrant, by MedChemExpress (1 mentions)
Mcf 7, by Merck Group (1 mentions)
Tamoxifen, by Merck Group (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Zr 75, by American Type Culture Collection (1 mentions)
2 protocols using ki67 antibody ab 16667
1
Tissue Immunohistochemistry Staining Protocol
2
Cell Line Authentication and Resistance Development
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MCF7, ZR75, and HEK293T cell lines were purchased from the American-Type Culture Collection (ATCC) and maintained in ATCC recommended medium. All model cells utilized were free of mycoplasma contamination and STR DNA profiling of the cells was used to confirm identity. MCF7-TamR cells were cultured in medium supplemented with 1 µmol/L of tamoxifen (H7904, Sigma, St. Louis, MO). MCF7-FR cells were cultured in medium supplemented with 1 µmol/L of fulvestrant (MedChem Express, Monmouth Junction, NJ) for 6 months to develop resistance. The SETDB1 antibody (11231-1-AP) was obtained from Proteintech (Rosemont, IL). The PELP1 antibody (A300-180A) was purchased from Bethyl Laboratories (Montgomery, TX). The ERα (04-820) and β-Actin antibodies (A-2066) were purchased from Millipore Sigma (Burlington, MA). The phospho-ER (Ser167) antibody (PA5-37570) was purchased from Invitrogen (Waltham, MA). The mCherry antibody (632543) was obtained from Takara Bio (San Jose, CA). The GFP antibody (632460) was obtained from Clontech. Antibodies for GST (2624), p-Akt (4060), Akt (9272), tri-methyl lysine motif (K-me3,14680) and GAPDH (8884) were purchased from Cell Signaling Technology (Beverly, MA). For IHC analysis, Ki67 antibody (ab16667) was purchased from Abcam (Cambridge, MA) and p-Akt (05-1003) antibody was purchased from Millipore Sigma.
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