Human Granzyme H HEK293 Overexpression Lysate: 产品信息
This Human Granzyme H overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Granzyme H protein (Cat: 10348-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human Granzyme H (NP_219491.1) (Met 1-Leu 246) with a C-terminal polyhistidine tag was expressed.
The secreted recombinant human Granzyme H comprises 239 amino acids with a predicted molecular mass of 26.7 kDa. As a result of glycosylation, rh Granzyme H migrates as an approximately 36 kDa band in SDS-PAGE under reducing conditions.
Human Granzyme H HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Granzyme H HEK293 Overexpression Lysate: 别称
Human CCP-X Overexpression Lysate; Human CGL-2 Overexpression Lysate; Human CSP-C Overexpression Lysate; Human CTLA1 Overexpression Lysate; Human CTSGL2 Overexpression Lysate; Human GZMH Overexpression Lysate
Granzyme H 背景信息
Granzymes are key components of the immune response that play important roles in eliminating host cells infected by intracellular pathogens. Several granzymes are potent inducers of cell death. A total of eight granzymes (A-G and M) have been identified in the mouse, but only five are known in humans (A, B, H, M, and granzyme 3), and granzyme H appears to be specifically human. Human granzyme H is a neutral serine protease that is expressed predominantly in the lymphokine-activated killer (LAK)/natural?killer (NK) compartment of the immune system. In adenovirus-infected cells in which granzyme B (gzmB) and downstream apoptosis pathways are inhibited, granzyme H directly cleaves the adenovirus DNA-binding protein (DBP), a viral component required for viral DNA replication. This virus demonstrated that gzmH directly induces an important decay in viral DNA replication. Interestingly, gzmH also cleaves the adenovirus 1K assembly protein, a major inhibitor of gzmB, and relieves gzmB inhibition. Granzyme H has a very high amino acid identity (>9%) with many portions of the granzyme B sequence, particularly near the amino terminus of the molecule despite performing a distinct enzymic function.
Meier M., et al.,(1990), Cloning of a gene that encodes a new member of the human cytotoxic cell protease family. Biochemistry 29:4042-4049.
Haddad P., et al., (1991), Structure and evolutionary origin of the human granzyme H gene.Int. Immunol. 3:57-66.
Klein J.L., et al.,(1990), Characterization of a novel, human cytotoxic lymphocyte-specific serine protease cDNA clone (CSP-C).Tissue Antigens 35:220-228.