Tumor tissue samples were collected from xenograft tumors, fixed in 10% neutral buffered formalin, paraffin-embedded, and cut into sections. The sections were heated in 10 mM citrate buffer (pH 6.0) for EGFR, HER2, and Ki67 or in EDTA buffer (pH 8.0) for HER3 with a Decloaking Chamber. Samples were incubated with rabbit anti-EGFR (Abcam; ab52894, 1:200), anti-HER2 (Cell Signaling; cs4290, 1:200), anti-HER3 (Cell Signaling; cs12708, 1:50), or anti-Ki67 (Cell Signaling; cs9027, 1:800). Sections were stained by VECTASTAIN universal kit/HRP (Vector Lab, Burlingame, CA). Antibody binding was revealed by addition of 3,3′-diaminobenzidine substrate. Tissues were counterstained with Mayer’s hematoxylin (Thermo Fisher Scientific, Waltham, MA) and were examined using an Olympus BX51 microscope.
Cs12708
Manufactured by Cell Signaling Technology
Cs12708 is a laboratory equipment product offered by Cell Signaling Technology. It is a core function piece of equipment used for scientific research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti her3, by Cell Signaling Technology (1 mentions)
Ab52894, by Cell Signaling Technology (1 mentions)
Vectastain universal kit hrp, by Vector Laboratories (1 mentions)
Mayer s hematoxylin, by Thermo Fisher Scientific (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
Matrigel solubilized basement membrane, by BD (1 mentions)
2 protocols using cs12708
1
Immunohistochemical Analysis of Xenograft Tumors
2
Xenograft Tumor Establishment and Immunohistochemistry
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All animal studies were conducted, and data collected with approval from the University of California Davis institutional animal care and use committee. Animal studies were performed as described previously (29 (link)). In summary, 4- to 5-week-old male Balb/c athymic Foxn1nu (nu/nu) mice (Harlan Sprague Dawley, Inc) were subcutaneously implanted in the flank with suspensions of CWR22 or 22Rv1 (2.5 million cells/site) in 50% Matrigel-solubilized basement membrane (BD Biosciences) and xenografts established. When palpable tumors were observed, the animals were castrated by bilateral scrotal excision, following isoflurane anesthetization or sham-operated by opening the animals surgically, but no tissues were removed. Mice were sacrificed when tumor size exceeded 150 mm in any one dimension or at the end of the study period. Mouse tumors were fixed in 10% buffered formalin (Medical Industries) for 30 min at RT, after which the pellet was immersed in 600-μl liquefied agar at 50 to 60 °C. The agar containing the tumor was paraffin-embedded and processed based on established protocols. Tumors were then stained with rabbit polyclonal anti-ErbB3 (CS-12708; 1:100 dilution, Cell Signaling Technologies).
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