BD PharmingenTM Purified Mouse Anti-C-Peptide

Description

Flow cytometric analysis of C-Peptide expression in human INS-transfected cells and a mouse insulinoma cell line. LEFT PANEL: Untransfected dashed-line histogram and human INS-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-C-Peptide Cat. No. 565830 followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . RIGHT PANEL: Beta-TC-6 cells ATCC CRL-11505 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were washed and stained with either Purified Mouse IgG1, kappa isotype control Cat. No. 554121, dashed-line histogram or Purified Mouse Anti-C-Peptide Cat. No. 565830, solid-line histogram followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . All fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto II flow cytometry system. LEFT PANEL: Western blot and peptide blocking analysis of C-Peptide expression. Lysate from INS-transfected 293F cells was prepared for electrophoresis SDS-PAGE in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes. The left membrane was probed with 8 lane 1 , 4 lane 2 , and 2 lane 3 mug/mL of Purified Mouse Anti-C-Peptide Cat. No. 565830 . The right membrane was probed with 4 mug/mL Purified Mouse Anti-C-Peptide lane 1 and with 4 mug/mL Purified Mouse Anti-C-Peptide that was mixed with 40 mug/mL C-Peptide for one hour lane 2 . Specific staining was detected with HRP Goat Anti-Mouse Ig Cat No 554002 . RIGHT PANEL: Immunohistochemical staining of C-Peptide in human, rat, and mouse islets of Langerhans. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , sections from formalin-fixed, paraffin-embedded human top row , rat middle row , and mouse bottom row pancreata were blocked using an Avidin/ Biotin Blocking Kit Vector Laboratories, Cat. No. SP-2001 as recommended by the manufacturer. The sections were then stained overnight with either Purified Mouse IgG1 kappa Isotype Control Cat. No. 550878, left column or Purified Mouse Anti-C-Peptide Cat. No. 565830, right column . A three-step staining procedure that employs either Biotin Goat Anti-Mouse Ig Cat. No. 550337, top and middle rows or Biotin Rat Anti-Mouse IgG1 Cat. No.553441, bottom row , Streptavidin HRP Cat. No. 550946 and DAB Cat. No. 550880 was used to reveal the primary staining reagents. Counterstaining was with Hematoxylin. Original magnifications: 40X. Flow cytometric analysis of C-Peptide expression in human INS-transfected cells and a mouse insulinoma cell line. LEFT PANEL: Untransfected dashed-line histogram and human INS-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-C-Peptide Cat. No. 565830 followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . RIGHT PANEL: Beta-TC-6 cells ATCC CRL-11505 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were washed and stained with either Purified Mouse IgG1, kappa isotype control Cat. No. 554121, dashed-line histogram or Purified Mouse Anti-C-Peptide Cat. No. 565830, solid-line histogram followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . All fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto II flow cytometry system. Show More LEFT PANEL: Western blot and peptide blocking analysis of C-Peptide expression. Lysate from INS-transfected 293F cells was prepared for electrophoresis SDS-PAGE in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes. The left membrane was probed with 8 lane 1 , 4 lane 2 , and 2 lane 3 mug/mL of Purified Mouse Anti-C-Peptide Cat. No. 565830 . The right membrane was probed with 4 mug/mL Purified Mouse Anti-C-Peptide lane 1 and with 4 mug/mL Purified Mouse Anti-C-Peptide that was mixed with 40 mug/mL C-Peptide for one hour lane 2 . Specific staining was detected with HRP Goat Anti-Mouse Ig Cat No 554002 . RIGHT PANEL: Immunohistochemical staining of C-Peptide in human, rat, and mouse islets of Langerhans. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , sections from formalin-fixed, paraffin-embedded human top row , rat middle row , and mouse bottom row pancreata were blocked using an Avidin/ Biotin Blocking Kit Vector Laboratories, Cat. No. SP-2001 as recommended by the manufacturer. The sections were then stained overnight with either Purified Mouse IgG1 kappa Isotype Control Cat. No. 550878, left column or Purified Mouse Anti-C-Peptide Cat. No. 565830, right column . A three-step staining procedure that employs either Biotin Goat Anti-Mouse Ig Cat. No. 550337, top and middle rows or Biotin Rat Anti-Mouse IgG1 Cat. No.553441, bottom row , Streptavidin HRP Cat. No. 550946 and DAB Cat. No. 550880 was used to reveal the primary staining reagents. Counterstaining was with Hematoxylin. Original magnifications: 40X. Show More Flow cytometric analysis of C-Peptide expression in human INS-transfected cells and a mouse insulinoma cell line. LEFT PANEL: Untransfected dashed-line histogram and human INS-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-C-Peptide Cat. No. 565830 followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . RIGHT PANEL: Beta-TC-6 cells ATCC CRL-11505 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were washed and stained with either Purified Mouse IgG1, kappa isotype control Cat. No. 554121, dashed-line histogram or Purified Mouse Anti-C-Peptide Cat. No. 565830, solid-line histogram followed by APC Goat Anti-Mouse Ig Cat. No. 550826 . All fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto II flow cytometry system. LEFT PANEL: Western blot and peptide blocking analysis of C-Peptide expression. Lysate from INS-transfected 293F cells was prepared for electrophoresis SDS-PAGE in a 2D Tris-Glycine polyacrylamide gel. The proteins were transferred to PVDF membranes. The left membrane was probed with 8 lane 1 , 4 lane 2 , and 2 lane 3 mug/mL of Purified Mouse Anti-C-Peptide Cat. No. 565830 . The right membrane was probed with 4 mug/mL Purified Mouse Anti-C-Peptide lane 1 and with 4 mug/mL Purified Mouse Anti-C-Peptide that was mixed with 40 mug/mL C-Peptide for one hour lane 2 . Specific staining was detected with HRP Goat Anti-Mouse Ig Cat No 554002 . RIGHT PANEL: Immunohistochemical staining of C-Peptide in human, rat, and mouse islets of Langerhans. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , sections from formalin-fixed, paraffin-embedded human top row , rat middle row , and mouse bottom row pancreata were blocked using an Avidin/ Biotin Blocking Kit Vector Laboratories, Cat. No. SP-2001 as recommended by the manufacturer. The sections were then stained overnight with either Purified Mouse IgG1 kappa Isotype Control Cat. No. 550878, left column or Purified Mouse Anti-C-Peptide Cat. No. 565830, right column . A three-step staining procedure that employs either Biotin Goat Anti-Mouse Ig Cat. No. 550337, top and middle rows or Biotin Rat Anti-Mouse IgG1 Cat. No.553441, bottom row , Streptavidin HRP Cat. No. 550946 and DAB Cat. No. 550880 was used to reveal the primary staining reagents. Counterstaining was with Hematoxylin. Original magnifications: 40X.