ELISA, Immunogen, SDS-PAGE, WB, Bioactivity testing in progress
Species
Homo sapiens (Human)
Nature
Recombinant
Endotoxin level
Please contact with the lab for this information.
Purity
>90% as determined by SDS-PAGE.
Predicted molecular weight
27.64 kDa
Form
Lyophilized
Storage buffer
Lyophilized from a solution in PBS pH 7.4, 0.02% NLS, 1 mM EDTA, 4% Trehalose, 1% Mannitol.
Please refer to the specific buffer information in the hardcopy of datasheet or the lot-specific COA.
Reconstitution
Reconstitute in sterile water for a stock solution. Adjust the volume of sterile water so that, after reconstitution, the final stock concentration matches the value indicated on the product label or datasheet.
RECONSTITUTION PROTOCOL
1. Allow the product to be reconstituted and the reconstitution diluent to reach room temperature.
2. Tap or briefly centrifuge the product vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.
3. Use the diluent and stock concentration recommended in the product datasheet. If preparing a stock concentration higher than recommended, contact Technical Service.
4. After adding the diluent, re-cap the vial and invert gently by hand or place on a slow rocking platform to allow the reconstitution diluent to coat all the surfaces inside the vial. Note: Do not mix by vortexing or by pipetting the material up and down.
5. Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.
6. Store the reconstituted material in single use aliquots in polypropylene or siliconized tubes. Aliquots of 10 μL or larger are recommended.
Shipping
In general, proteins are provided as lyophilized powder/frozen liquid. They are shipped out with dry ice/blue ice unless customers require otherwise.
Stability and Storage
Use a manual defrost freezer and avoid repeated freeze thaw cycles. Store at 2 to 8°C for frequent use. Store at -20 to -80°C for twelve months from the date of receipt.
Granzyme B (GZMB) is a ~27 kDa protein. Abundant protease in the cytosolic granules of cytotoxic T-cells and NK-cells which activates caspase-independent pyroptosis when delivered into the target cell through the immunological synapse. It cleaves after Asp. Once delivered into the target cell, acts by catalyzing cleavage of gasdermin-E (GSDME), releasing the pore-forming moiety of GSDME, thereby triggering pyroptosis and target cell death. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -9 and -10 (CASP3, CASP9 and CASP10, respectively) to give rise to active enzymes mediating apoptosis.
1. Poe, M. et al. (1991) The Journal of biological chemistry 266, 98-103. PMID: 1985927
2. Hameed, A. et al. (1988) Journal of immunology (Baltimore, Md. : 1950) 141, 3142-7. PMID: 3262682
3. Krähenbühl, O. et al. (1988) Journal of immunology (Baltimore, Md. : 1950) 141, 3471-7. PMID: 3263427
4. Froelich, CJ. et al. (1993) Journal of immunology (Baltimore, Md. : 1950) 151, 7161-71. PMID: 8258716
5. Liu, Y. et al. (2020) Science immunology 5. PMID: 31953257
6. Zhang, Z. et al. (2020) Nature 579, 415-420. PMID: 32188940
7. Yang, X. et al. (1998) The Journal of biological chemistry 273, 34278-83. PMID: 9852092
Note
For research use only.
Images
SDS-PAGE
SDS-PAGE for Recombinant Human GZMB Protein, N-His
Western blot
Detected by Anti-GZMB monoclonal antibody (Catalog No: HY574013)
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