Anti-Grp75/MOT antibody [JG1]

10 product images

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  Flow Cytometry - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Overlay histogram showing HepG2 cells stained with ab2799 (red line). The cells were fixed with 80% methanol (5 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 (ab2799, 1/100 dilution) for 30 min at 22ºC. 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 22ºC. Isotype control antibody (black line) was mouse IgG3 [MG3-35] (Mouse IgG3, kappa monoclonal [MG3-35] - Isotype Control ab18394, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Grp75/MOT (green) with ab2799. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were incubated with (left panel) or without (right panel) ab2799 (1:50) for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody (1:400) for 30 minutes at room temperature. F-Actin (red) was stained with Dylight 554 phalloidin, and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunohistochemistry was performed on normal biopsies of deparaffinized human testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a Mouse Monoclonal Antibody recognizing Grp75/MOT (ab2799) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  ICC/IF image of ab2799 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2799, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/ Immunofluorescence of P19 (Mouse embryonal carcinoma cell line) cells using ab2799.

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/ Immunofluorescence of NS-1 (Mouse myeloma cell line) cells using ab2799.

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/ Immunofluorescence of HMVEC (Human microvascular endothelial cell line) cells using ab2799.

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/ Immunofluorescence of A549 (Human lung carcinoma cell line) cells using ab2799.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [JG1] (ab2799)

  ab2799 (1μg/ml) staining GrP75/MOT in human frontal cerebral cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is strong mitochondrial staining.
  Insert depicts negative control (no primary antibody).
  Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

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  Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [JG1] (ab2799)

  Immunocytochemistry/Immunofluorescence analysis of human fibroblasts labelling Grp75/MOT with ab2799.