IHC staining was performed as described previously 12 (link). In brief, formalin-fixed paraffin-embedded sections were de-paraffinized, rehydrated and washed in Phosphate Buffer solution. After antigen retrieval in 10 mM sodium citrate buffer, blocking of peroxidase activity with 3% H2O2, and non-specific protein blocking using 10% goat seum, sections were incubated with primary antibodies (anti-MCT1 at a dilution of 1:100, Abcam, Cambridge, MA, USA), followed by anti-mouse/rabbit horseradish peroxidase-labeled antibody (Univ-bio, Shanghai, China) as the second antibody. Membranous and cytoplasmic MCT1 expression was observed, and the evaluation of MCT1 staining was defined considering the sum score of the proportion of positively stained cells (0, 0% of positive cells; 1, <5% of positive cells; 2, 5-50% positive cells; 3, >50% of positive cells) and the intensity of the staining (0, negative; 1, weak; 2, intermediate; 3, strong) as described previously 13 (link). MCT1 staining was scored as 0 (sum score 0), 1 (sum score 1, 2), 2 (sum score 3, 4), and 3 (sum score 5, 6). Score of 0 and 1 were considered as low expression, while 2 and 3 were as high expression.
Anti mct1
Manufactured by Abcam
Sourced in United States
Anti-MCT1 is a lab equipment product that is used to detect the presence and expression levels of the MCT1 (Monocarboxylate Transporter 1) protein. MCT1 is a membrane transport protein involved in the movement of various monocarboxylates, such as lactate and pyruvate, across cell membranes. This product can be utilized in research applications to study the role of MCT1 in cellular processes and metabolic pathways.
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Lab products found in correlation
2 protocols using anti mct1
1
Immunohistochemical Evaluation of MCT1 Expression
2
Postnatal Expression Changes in Rat Corpus Callosum
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To study the expression change pattern of MCT1, MBP, GFAP and vWF in postnatal rat corpus callosum, the expression levels were quantified by Western blotting. Tissue samples were micro-dissected and homogenized in RIPA lysis buffer with protease inhibitor cocktail which containing 0.1 % Nonidet P-40, 1 mM dithiothreitol, 10 mM EDTA, 40 mM Tris-HCl (pH 7.4), 120 mM NaCl for 30 min on ice to promote lysis, and then spun down at 12,000 rpm for 10 min at 4 ºC. Protein concentrations were quantified using BCA assay (Beyotime Institute of Biotechnology, China). Equal amounts of protein (20 μg/lane) were separated on 10 % SDS-PAGE gels, and the protein was transferred to nitrocellulose membranes. Membranes were blocked with 5 % non-fat milk in PBS for 1 h and then incubated overnight at 4 ºC with the following primary antibodies: anti-MCT1 (abcam 1:1,000), anti-MBP (chemicon 1:800), anti-GFAP (sigma 1:2,000) and anti-vWF (abcam 1:1,000). After three washes with Tris-Tween buffered saline, membranes were subsequently incubated with fluorescent secondary antibodies (IRDye 700 or IRDye 800) for 2 h at R/T on a shaker. Lastly, membranes were detected by Odyssey Infrared Imaging System (LI-CON, Biotechnology Inc., Lincoln, USA). Band size and density measurements from each sample were collected using ImageJ. Values were normalized by the levels of β-actin.
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