domestic rabbit monoclonal (PSH04-06)
reactivity: human
application: western blot, flow cytometry
reactivity: human
application: western blot, flow cytometry
Western blot analysis of GITR on HUT 102 cell lysates with Rabbit anti-GITR antibody (HA722087) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 26 kDa Observed band size: 26 kDa Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722087) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HUT 102 cells labeling GITR with Rabbit anti-GITR antibody (HA722087) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GITR antibody (HA722087) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Flow cytometric analysis of HUT 102 cells labeling GITR. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722087, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
quantity: 100μl
price: 385.00 USD
to the supplier
domestic rabbit monoclonal (PSH05-63)
reactivity: human
application: ELISA
reactivity: human
application: ELISA
Sandwich ELISA analysis of human GITR matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722383) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted recombinant human GITR protein (HA210945) starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA722384, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Interpolated concentrations of native GITR in Jurkat and HuT-102 cell extract samples based on a 1000 µg/ml extract load. The concentrations of GITR were interpolated from the GITR standard curve and corrected for sample dilution. The mean GITR concentration was determined to be 37.69 ng/ml in HuT-102 cell extract. There was no detectable signal in Jurkat cell extract.
Interpolated concentrations of spiked GITR in human cell culture media samples. The concentrations of GITR were measured in duplicates, interpolated from the GITR standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.
quantity: 100μl
price: 649.00 USD
to the supplier
domestic rabbit monoclonal (PSH05-64)
reactivity: human
application: ELISA
reactivity: human
application: ELISA
Sandwich ELISA analysis of human GITR matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722383) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted recombinant human GITR protein (HA210945) starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA722384, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Interpolated concentrations of native GITR in Jurkat and HuT-102 cell extract samples based on a 1000 µg/ml extract load. The concentrations of GITR were interpolated from the GITR standard curve and corrected for sample dilution. The mean GITR concentration was determined to be 37.69 ng/ml in HuT-102 cell extract. There was no detectable signal in Jurkat cell extract.
Interpolated concentrations of spiked GITR in human cell culture media samples. The concentrations of GITR were measured in duplicates, interpolated from the GITR standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.
quantity: 100μl
price: 649.00 USD
to the supplier
domestic rabbit monoclonal (PSH05-64)
reactivity: human
conjugate: biotin
application: ELISA
reactivity: human
conjugate: biotin
application: ELISA
domestic rabbit monoclonal (PSH04-06)
reactivity: human
application: western blot, flow cytometry
reactivity: human
application: western blot, flow cytometry
Western blot analysis of GITR on HUT 102 cell lysates with Rabbit anti-GITR antibody (HA750929) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 26 kDa Observed band size: 26 kDa Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750929) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HUT 102 cells labeling GITR with Rabbit anti-GITR antibody (HA750929) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GITR antibody (HA750929) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Flow cytometric analysis of HUT 102 cells labeling GITR. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA750929, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
quantity: 100μl
price: 649.00 USD
to the supplier