BD Pharmingen™ Purified Mouse Anti-MLC2v

Flow cytometric analysis of MLC2v and MLC2a expression in MLC2v-transfected 293F cells. Untransfected (dashed line histogram) and human MLC2v-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Purified Mouse Anti-Human MLC2v (Cat. No. 565496, left panel) and Purified Mouse Anti-Human MLC2a (Cat. No. 565496, middle panel) followed by PE Goat Anti-Mouse Ig (Cat. No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.

Western blot analysis of MLC2v expression (right panel). Lysate from MLC2v-transfected 293F cells was prepared for electrophoresis (SDS-PAGE) in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes and then probed with 0.063 (lane 1), 0.032 (lane 2), and 0.016 (lane 3) µg/mL of Purified Mouse Anti-MLC2v. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat No 554002).

Immunohistochemical staining of MLC2v in human left ventricle and left atrium. Following antigen retrieval with BD Retrievagen A buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 550878 ; top left) or Purified Mouse Anti-MLC2v (top right) on left ventricle. Additionally, Purified Mouse Anti-MLC2v (bottom left) and Purified Mouse Anti-MLC2a (Cat. No. 565496, bottom right) were used to stain left atrium. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Ig (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissues were counterstained with Hematoxylin. Original magnification: 20X (MLC2v) and 40X (MLC2a).