Anti-ENPP3 antibody (bs-1568R)

Anti-ENPP3 antibody

DATASHEET

Host:Rabbit

Target Protein:ENPP3

IR:Immunogen Range:40-140/875

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:5169

Swiss Prot:O14638

Source:KLH conjugated synthetic peptide derived from human ENPP3:40-140/875 

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:The protein encoded by this gene belongs to a series of ectoenzymes that are involved in hydrolysis of extracellular nucleotides. These ectoenzymes possess ATPase and ATP pyrophosphatase activities and are type II transmembrane proteins. Expression of the related rat mRNA has been found in a subset of immature glial cells and in the alimentary tract. The corresponding rat protein has been detected in the pancreas, small intestine, colon, and liver. The human mRNA is expressed in glioma cells, prostate, and uterus. Expression of the human protein has been detected in uterus, basophils, and mast cells.

Size:100ul

Concentration:1mg/ml

Applications:ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(2μg /test)
IF(1:200-800)

Cross Reactive Species:Human
Mouse
Rat
Dog
Pig
Cow
.

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

VALIDATION IMAGES

Paraformaldehyde-fixed, paraffin embedded (Rat small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation: Anti-ENPP3 Antibody, conjugated (bs-1568R-A488) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (Rat small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation: Anti-ENPP3 Antibody, conjugated (bs-1568R-A488) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.

Blank control: Raji (blue).
Primary Antibody: Rabbit Anti-ENPP3 antibody(bs-1568R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange),used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (bs-1568R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

Black line : Positive blank control (Raji); Negative blank control (Molt4）
Green line : Primary Antibody (Rabbit Anti-ENPP3 antibody (bs-1568R) )
Orange line：Isotype Control Antibody (Rabbit IgG) .
Blue line : Secondary Antibody (Goat anti-rabbit IgG-PE)/Goat anti-rabbit IgG-AF488)
Raji（Positive）and Molt4（Negative control）cells (black) were incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ENPP3 Antibody(bs-1568R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).