Usage
Western Blot, Flow Cytometry, Immunohistochemistry, Direct ELISA. Western Blot: This antibody can be used at 1-2 ug/mL with the appropriate secondary reagents to detect human Granzyme B. Using a colorimetric detection system, the detection limit for rhGranzyme B is approximately 5 ng/lane under non-reducing and reducing conditions. Chemiluminescent detection will increase sensitivity by 5 to 50 fold. Flow Cytometry: This antibody was tested for flow cytometry using NK-92 cells. For intracellular staining to detect human Granzyme B, cells must first be fixed and permeabilized using 4% paraformaldehyde and 0.1% saponin in PBS. Dilute this antibody to 25 ug/mL and add 10 ul of the diluted solution to 1-5 x 10^5 cells in a total reaction volume not exceeding 200ul. Following a 30 minute incubation, cells should be washed with 0.1% saponin prior to addition of a secondary developing reagent. The binding of unlabeled monoclonal antibodies may be visualized by adding 10 ul of a 25 ug/mL solution of a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome. Cells should be washed for a final time in 0.1% saponin prior to flow cytometric analysis. Immunohistochemistry: This antibody was used at a concentration of 8-25 ug/mL with appropriate secondary reagents to detect human Granzyme B in paraformaldehyde-fixed human PBMC. For chromogenic detection of labeling, the use of Cell and Tissue Staining Kits is recommended. Direct ELISA: This antibody can be used at 0.5-1.0 ug/mL with the appropriate secondary reagents to detect human Granzyme B. The detection limit for rhGranzyme B is approximately 5 ng/well.
Presentation
Lyophilized from PBS, 5% Trehalose
Reconstitution
Reconstitute in 200 ul sterile PBS
Storage
Lyophilized and resuspended forms: store at -20°C.
Restrictions
For research use only. Intended for use by laboratory professionals.
Guarantee
This antibody carries the LSBio 100% Guarantee.
LSBio Guarantee