primary antibodies against mouse b7-2 (cd86)  (Santa Cruz Biotechnology)

Journal: Experimental Animals

Article Title: Moderate hyperglycemia suppresses melanoma metastasis to liver

doi: 10.1538/expanim.22-0078

Figure Lengend Snippet: Effects of moderate hyperglycemia on the expression of TNFα , Cxcl10 , and macrophage markers in the liver. (A) TNFα expression. (B) Cxcl10 expression. (C–F) macrophage markers. Every gene expression intensity was normalized to that of GAPDH . Bars are mean ± SEM, n=3 ( TNFα ), n=6 ( Cxcl10 , F4/80 , CD68 , CD86 , CD206 ). (G) Protein band of CD86 and GAPDH of Wild and GKKO mice liver samples (a) and their intensity profiles analyzed using ImageJ (b). Blue dotted line: Wild, red dotted line: GKKO.

Article Snippet: The PVDF membrane was then blocked in TBS-T (Tris-buffered saline (25 mM Tris, pH 7.4, 150 mM NaCl) containing 1 (v/v)% Tween 20) solution containing 5 (w/v)% skim milk at 25°C for 30 min. Then the PVDF membrane was probed with primary antibodies against mouse B7-2 (CD86) (1:500, sc-28347; Santa Cruz Biotechnology, Dallas, TX, USA), and GAPDH (1:1,000, sc-32233; Santa Cruz Biotechnology) in a 5% skim milk TBS-T solution at 25°C for 3 h. The membrane was further incubated with secondary antibody (anti-mouse immunoglobulin conjugated to alkaline phosphatase, Promega, Madison, WI, USA) in TBS-T at 25°C for 1 h. Finally, membranes were incubated with Western Blue Stabilized Substrate (Promega) for alkaline phosphatase at 25°C for 10 min.

Techniques: Expressing, Gene Expression