Active Recombinant Human GZMB Protein (19-247aa), C-His tagged 

Cat.No. :GZMB-33H

Product Overview :Recombinant human GZMB (19-247aa), fused to His-tag at C-terminus, was expressed in insect cell and purified by using conventional chromatography techniques.

Species :Human

Source :Insect Cells

Tag :His

Protein Length :19-247 aa

Description :This gene encodes a member of the granzyme subfamily of proteins, part of the peptidase S1 family of serine proteases. The encoded preproprotein is secreted by natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) and proteolytically processed to generate the active protease, which induces target cell apoptosis. This protein also processes cytokines and degrades extracellular matrix proteins, and these roles are implicated in chronic inflammation and wound healing. Expression of this gene may be elevated in human patients with cardiac fibrosis.

Form :Liquid

Bio-activity :> 7, 000 pmol/min/μg, and is defined as the amount of enzyme that cleave 1pmole of Boc-Ala-Ala-Asp-SBzl at 37 centigrade.

Molecular Mass :26.5 kDa (235 aa)

AA Sequence :GEIIGGHEAKPHSRPYMAYLMIWDQKSLKRCGGFLIRDDFVLTAAHCWGSSINVTLGAHNIKEQEPTQQFIPVKRPIPHPAYNPKNFSNDIMLLQLERKAKRTRAVQPLRLPSNKAQVKPGQTCSVAGWGQTAPLGKHSHTLQEVKMTVQEDRKCESDLRHYYDSTIELCVGDPEIKKTSFKGDSGGPLVCNKVAQGIVSYGRNNGMPPRACTKVSSFVHWIKKTMKRY< HHHHHH>

Endotoxin :< 1 EU/μg of protein (determined by LAL method)

Purity : > 95 % by SDS-PAGE

Applications :SDS-PAGE, Enzyme Activity

Storage :Can be stored at +2 to +8 centigrade for 1 week. For long term storage, aliquot and store at -20 to -80 centigrade. Avoid repeated freezing and thawing cycles.

Concentration : 0.5 mg/mL (determined by absorbance at 280nm)

Storage Buffer :Phosphate-Buffered Saline (pH 7.4) containing 20 % glycerol, 1mM DTT.

Gene Name:GZMB granzyme B [ Homo sapiens (human) ]

Official Symbol:GZMB

Synonyms:GZMB; granzyme B; C11; HLP; CCPI; CGL1; CSPB; SECT; CGL-1; CSP-B; CTLA1; CTSGL1; granzyme B; T-cell serine protease 1-3E; cathepsin G-like 1; cytotoxic T-lymphocyte proteinase 2; cytotoxic serine protease B; fragmentin 2; granzyme B (granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1); human lymphocyte protein; EC 3.4.21.79

GENE INFORMATION:

Gene ID:3002

mRNA Refseq:NM_004131

Protein Refseq:NP_004122

MIM:123910

UniProt ID:P10144

Active Recombinant Human PRF1 protein, His-GST-tagged

Cat.No. :PRF1-25H

Product Overview :Recombinant Human PRF1 protein(Lys32-Phe316), fused with N-terminal His and GST tag, was expressed in E. coli.

SPECIFICATIONS

Species :Human

Source :E. coli

Tag :GST&His

Protein Length :32-316 aa

Form :20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300.

Bio-activity :1. Perforin 1 (PRF1) is a pore forming cytolytic protein found in the granules of cytotoxic T lymphocytes (CTLs) and NK cells. Upon degranulation, perforin binds to the target cell's plasma membrane, and oligomerises in a Ca2 dependent manner to form pores on the target cell. The pore formed allows for the passive diffusion of a family of pro-apoptotic proteases, known as the granzymes, into the target cell. Besides, Calreticulin (CRT) has been identified as an interactor of PRF1, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PRF1 and recombinant human CRT. Briefly, PRF1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to CRT-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-PRF1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of PRF1 and CRT was shown in Figure 2, and this effect was in a dose dependent manner.2. The activity of recombinant PRF1 was measured by lysis of erythrocytes using a hemolysis assay. A general procedure is as fllows: two-fold dilute the recombinant human PRF1 with 0.9% NaCl, add 50μL a serial dilution of PRF1, 10μL 0.1M CaCl2 to each well, then add 50μL 0.25% rabbit erythrocyte (RaE) to each well and mixed gently. Add 10μL 0.9% NaCl to reaplace CaCl2 in control wells. The plate is incubated for 20 hours at 37°C, 5% CO2. The results are shown in Figure 3. It was obvious that the minimal effective concentration of PRF1 is 12.5μg/mL.

Molecular Mass :62kDa

Purity : > 95%

Storage :Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

Reconstitution :Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

GENE INFORMATION

Gene Name:PRF1 perforin 1 (pore forming protein) [ Homo sapiens ]

Official Symbol:PRF1

Synonyms:PRF1; perforin 1 (pore forming protein); perforin-1; HPLH2; P1; Perforin; perforin 1 (preforming protein); PFP; cytolysin; lymphocyte pore forming protein; lymphocyte pore-forming protein; FLH2; PFN1; MGC65093

Gene ID:5551

mRNA Refseq:NM_001083116

Protein Refseq:NP_001076585

MIM:170280

UniProt ID:P14222

Active Recombinant Full Length Human PRF1 protein

Cat.No. :PRF1-158H

Product Overview :Recombinant Human PRF1 protein(Pro22~Trp555), fused to N-terminal His tag, was expressed in E. coli.

SPECIFICATION

Species :Human

Source :E.coli

Tag :His

Protein Length :22-555 a.a.

Form :20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl and 5% trehalose

Molecular Mass :63kDa as determined by SDS-PAGE reducing conditions.

Endotoxin :< 1.0 EU per μg protein as determined by the LAL method.

Purity : > 80 %

Storage :Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

Reconstitution :Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex.

GENE INFORMATION

Gene Name:PRF1 perforin 1 (pore forming protein) [ Homo sapiens ]

Official Symbol:PRF1

Synonyms:PRF1; perforin 1 (pore forming protein); perforin-1; HPLH2; P1; Perforin; perforin 1 (preforming protein); PFP; cytolysin; lymphocyte pore forming protein; lymphocyte pore-forming protein; FLH2; PFN1; MGC65093;

Gene ID:5551

mRNA Refseq:NM_001083116

Protein Refseq:NP_001076585

MIM:170280

UniProt ID:P14222