Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control

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  Flow Cytometry - Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

  Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with Anti-CD82 antibody [TS82b] - BSA and Azide free ab59509 (red line).
  

  The cells were fixed with 80% methanol (5 minutes) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-CD82 antibody [TS82b] - BSA and Azide free ab59509, 1 μg/1x10⁶ cells) for 30 minutes at 22°C. The secondary antibody used was a goat anti-mouse DyLight^® 488 (IgG, H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 minutes at 22°C.

  Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 μg/1x10⁶ cells) used under the same conditions.

  Acquisition of >5,000 events was performed.

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  Hirakawa et al PLoS One. 2016 Aug 3;11(8):e0159912. doi: 10.1371/journal.pone.0159912. eCollection 2016. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Flow Cytometry - Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

  Effects of hypoxia on mRNA expression of IGF1R and IGF1, and production of IGF1 in cancer cells.

  Cell surface expression of IGF1R by FACScan analysis. IGF1R expression level of Panc-1, RWP-1, OCUP-AT, and MiaPaCa-2 cells was higher in hypoxia than that in normoxia.

  Cells (2 x 10⁶ cells/mL) were fixed with 2% paraformaldehyde and incubated in PBS with anti IGF1R antibody (Anti-IGF1 Receptor antibody [alphaIR3] ab16890, Abcam) or mouse IgG1- isotype control (ab91353, Abcam) for 30 minutes at 22°C. Cells were subsequently labeled with FITC-conjugated secondary antibody (1:500; Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879, Abcam) for 30 minutes at 22°C.

  The percentage of positive cells were calculated and compared with isotype-matched control-stained cells.

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  Karanovic et al One. 2016 Aug 25;11(8):e0161706. doi: 10.1371/journal.pone.0161706. eCollection 2016. Fig 9. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

  Glomerular nestin expression in experimental groups.

  (A) Negative control (×200).

  (C) SHADR group: Diffuse glomerular nestin expression was detected involving almost all podocytes within glomerulus. After losartan and tempol treatment, either single or in combination, kidneys restored nestin expression similar to controls (SHC group).

  Immunostaining was applied on 5 μm thick paraffin sections. After deparaffinization and rehydration, the sections were treated by microwave for 20 minutes at 400 W in citrate buffer (pH 6.0). After antigen retrieval, samples were incubated for 1 hour at room temperature with primary antibody for nestin (dilution 1:100). Sections were then treated using 3-amino-9-ethylcarbazole (AEC) as substrate, and counterstained with hematoxylin. Negative controls were performed by omitting the first antibody and mouse monoclonal antibodies as isotype control mouse IgG1 (ab91353) antibody was also used.

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  Flow Cytometry (Intracellular) - Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)