Anti-CD105 antibody [MEM-226]

6 product images

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  Flow Cytometry - Anti-CD105 antibody [MEM-226] (ab2529)

  Overlay histogram showing U937 (Human histiocytic lymphoma cell line) cells stained with ab2529 (red line).
  

  The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2529, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in U937 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  

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  Western blot - Anti-CD105 antibody [MEM-226] (ab2529)

  Lanes 1 - 3: Merged signal (red and green). Green - ab2529 observed at 70 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
  

  ab2529 was shown to recognize ENG (Endoglin) in wild-type HeLa cells as signal was lost at the expected MW in ENG knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ENG knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab2529 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-CD105 antibody [MEM-226] (ab2529) at 1/1000 dilution

  Lane 1: Wild-type HeLa whole cell lysate at 20 µg

  Lane 2: CD105 knockout HeLa whole cell lysate at 20 µg

  Lane 3: HUVEC whole cell lysate at 20 µg

  Predicted band size: 70 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-CD105 antibody [MEM-226] (ab2529)

  ICC/IF image of ab2529 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
  

  The cells were 4% formaldehyde fixed (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2529, 10 μg/ml) overnight at +4°C. The secondary antibody (green) was ab69879, DyLight^® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 μM.

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  Sandwich ELISA - Anti-CD105 antibody [MEM-226] (ab2529)

  Standard Curve for CD105 (Analyte: CD105 protein (Recombinant human CD105 protein ab54338)); dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [MEM-226] to CD105 (ab2529) at 5ug/ml and Detector Antibody Rabbit polyclonal to CD105 (Anti-CD105 antibody ab21224) at 0.5ug/ml.

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  Western blot - Anti-CD105 antibody [MEM-226] (ab2529)

  All lanes: Western blot - Anti-CD105 antibody [MEM-226] (ab2529) at 5 µg/mL

  All lanes: Human colon tissue lysate at 10 µg

  Secondary

  All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Predicted band size: 70 kDa

  Observed band size: 45 kDa, 55 kDa, 80 kDa

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  Flow Cytometry - Anti-CD105 antibody [MEM-226] (ab2529)

  Flow cytometry analysis showing separation of HUVEC cells stained using ab2529 (concentration in sample 1.67 ?g/ml, GAM APC, red-filled) from HUVEC cells unstained by primary antibody (GAM APC, black-dashed).