Anti-Neutrophil Elastase antibody (bs-6982R)

Anti-Neutrophil Elastase antibody

DATASHEET

Host:Rabbit

Target Protein:Neutrophil Elastase

IR:Immunogen Range:101-200/267

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:1991

Swiss Prot:P08246

Source:KLH conjugated synthetic peptide derived from human Neutrophil Elastase/ELANE:101-200/267 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode the structurally similar proteins. The product of this gene hydrolyzes proteins within specialized neutrophil lysosomes, called azurophil granules, as well as proteins of the extracellular matrix following the protein's release from activated neutrophils. The enzyme may play a role in degenerative and inflammatory diseases by its proteolysis of collagen-IV and elastin of the extracellular matrix. This protein degrades the outer membrane protein A (OmpA) of E. coli as well as the virulence factors of such bacteria as Shigella, Salmonella and Yersinia. Mutations in this gene are associated with cyclic neutropenia and severe congenital neutropenia (SCN). This gene is clustered with other serine protease gene family members, azurocidin 1 and proteinase 3 genes, at chromosome 19pter. All 3 genes are expressed coordinately and their protein products are packaged together into azurophil granules during neutrophil differentiation. [provided by RefSeq, May 2009].

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
Flow-Cyt(1μg/Test)

Cross Reactive Species:Human
Mouse
Rat
Dog
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
Spinal cord (Mouse) Lysate at 40 ug
Primary: Anti-Neutrophil Elastase (bs-6982R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 48 kD

Sample:
HL60(Human) Cell Lysate at 30 ug
Primary: Anti-Neutrophil Elastase (bs-6982R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 41 kD

Sample:
Cerebrum (Mouse) Lysate at 40 ug
Primary: Anti-Neutrophil Elastase (bs-6982R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 31/46 kD

Blank control: U937.
Primary Antibody (green line): Rabbit Anti-CD30 antibody (bs-6982R-AF488)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control Black line): A549
Primary Antibody (orange line): Rabbit Anti-Neutrophil Elastase/AF488 Conjugated antibody (bs-6982R-AF488)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG-AF488(CST).
Protocol
The cells were fixed with 4% paraformaldehyde (10 min) , then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions followed by the antibody for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature.Acquisition of 20,000 events was performed.

Blank control: A549(blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice..
Isotype Control Antibody: Rabbit IgG(orange) ; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:100 in 1 X PBS containing 0.5% BSA ; Primary Antibody Dilution: 1μl in 100 μL1X PBS containing 0.5% BSA(green).