Following deparaffinization and rehydration, tumor sections (3-μm thickness) were incubated in 0.3% H2O2 in methanol for 30 min at 37 °C to block endogenous peroxidase. The sections were then boiled in 10 mmol/L citrate buffer (pH 6.0) for 2 min in an autoclave. The anti-ASIC2 (Abcam) or anti-NFAT1 (Cell Signaling) antibody was added and the sections were incubated at 4 °C overnight. The sections were visualized by using the diaminobenzidine solution (DAKO, Carpinteria, CA, USA), and then lightly counterstained with hematoxylin. Sections without incubation with primary antibody served as negative controls. The intensity of staining (brown color) was semi-quantitatively scored as follows: 1, weak; 2, medium; 3, strong; and 4, very strong. The percentage of maximally stained tumor cells in each section was recorded (0, <5%; 1, 5–30%; 2, 30–50%; 3, >50%). High expression of ASIC2 was defined as a combined score for the intensity and area of staining that was larger than 4. The results were verified by two pathologists independently.
Anti asic2
Manufactured by Abcam
Sourced in United States
Anti-ASIC2 is a primary antibody that specifically binds to the ASIC2 protein. ASIC2 is a subunit of the acid-sensing ion channel. This antibody can be used for the detection of ASIC2 in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Diaminobenzidine solution, by Agilent Technologies (1 mentions)
Anti nfat1, by Cell Signaling Technology (1 mentions)
Polyvinyl difluoride membrane, by Merck Group (1 mentions)
Anti na k atpase, by Abcam (1 mentions)
Anti vegf, by Bioss Antibodies (1 mentions)
Anti asic3, by Abcam (1 mentions)
Image lab, by Bio-Rad (1 mentions)
Anti β actin, by ZSGB-BIO (1 mentions)
Total protein extraction kit, by BestBio (1 mentions)
2 protocols using anti asic2
1
Immunohistochemical Quantification of ASIC2 and NFAT1
2
Western Blot Analysis of Ion Channels
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Cell culture total protein was extracted using a total protein extraction kit (Bestbio, Shanghai, China), protein samples were separated using a 10% SDS-polyacrylamide gel, and then transferred to a polyvinyl difluoride membrane (Millipore, USA). The membrane was then blocked with TBST (10 mM Tris, 150 mM NaCl, and 0.05% Tween 20 (pH 8.3)) containing 5% skim milk for 1 h at room temperature. Membranes were incubated overnight at 4 °C with anti-ASIC1a (Affinity, Cell Signal Transduction, USA), anti-ASIC2 (Abcam, Cambridge, UK), anti-ASIC3 (Abcam, Cambridge, UK), anti-ASIC4 (Abcam, Cambridge, UK), anti-Na + /K + -ATPase (Abcam, Cambridge, UK), anti-VEGF (Bioss, Beijing, China), and anti-β-actin (Zsbio, Beijing, China) antibodies. Membranes were washed in TBST and incubated with the corresponding secondary antibody (1:10000) for 2 h at room temperature. The signal was observed with a chemiluminescence imager (Bio-RAD, USA) according to the manufacturer's instructions. Images were quantified using the software Image lab (Bio-RAD, USA).
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