Anti-phospho-IRS1 (Ser323) antibody (bs-12709R)

Anti-phospho-IRS1 (Ser323) antibody

DATASHEET

Host:Rabbit

Target Protein:phospho-IRS1 (Ser323)

IR:Immunogen Range:PG(p-S)FR

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:3667

Swiss Prot:P35568

Source:KLH conjugated synthesised phosphopeptide derived from human IRS1 around the phosphorylation site of Ser323:PG(p-S)FR 

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:This gene encodes a protein which is phosphorylated by insulin receptor tyrosine kinase. Mutations in this gene are associated with type II diabetes and susceptibility to insulin resistance. [provided by RefSeq, Nov 2009]

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test)
ICC(1:100-500)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Rabbit
.

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

VALIDATION IMAGES

Sample:Ovary (Mouse) Lysate at 40 ug
Primary: Anti-phospho-IRS1 (Ser323)(bs-12709R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 132kD
Observed band size: 132kD

Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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; Antibody incubation with (phospho-IRS1 (Ser323)) Polyclonal Antibody, Unconjugated (bs-12709R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-phospho-IRS1 (Ser323) antibody (bs-12709R),Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE,Dilution: 1μg /test.

Blank control: HepG2.
Primary Antibody (green line): Rabbit Anti-phospho-IRS1 (Ser323) antibody (bs-12709R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.