phospho-IRS1 (Ser616) Polyclonal Antibody

$HepG2 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with IRS1 (Ser616) Polyclonal Antibody(bs-12710R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

HepG2 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with IRS1 (Ser616) Polyclonal Antibody(bs-12710R)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 incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Paraformaldehyde-fixed, paraffin embedded Rat uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with IRS1 (Ser616) Polyclonal Antibody, Unconjugated (bs-12710R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat uterus; 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\u00b0C for 30min; Antibody incubation with IRS1 (Ser616) Polyclonal Antibody, Unconjugated (bs-12710R) at 1:200 overnight at 4\u00b0C, DAB staining.

$A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-IRS1 (Ser616)) polyclonal Antibody, Unconjugated (bs-12710R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-IRS1 (Ser616)) polyclonal Antibody, Unconjugated (bs-12710R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

$Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Mouse Cerebellum tissue lysates probed with phospho-IRS1 (Ser616) Polyclonal Antibody, Unconjugated (bs-12710R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Mouse Cerebellum tissue lysates probed with phospho-IRS1 (Ser616) Polyclonal Antibody, Unconjugated (bs-12710R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Sheep
Pig
Horse
Chicken
Rabbit

Overview
Catalog #	bs-12710R
Product Name	phospho-IRS1 (Ser616) Polyclonal Antibody
Applications	WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Sheep, Pig, Horse, Chicken, Rabbit
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthesised phosphopeptide derived from human IRS1 around the phosphorylation site of Ser616
Modification Site	Ser616
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3667
Swiss Prot	P35568
Subcellular location	Cytoplasm, Nucleus, Cell membrane
Synonyms	IRS1 (phospho S616); p-IRS1 (phospho S616); HIRS 1; HIRS1; Insulin Receptor Substrate 1; IRS 1; IRS-1; IRS1; IRS1_HUMAN; OTTHUMP00000164234
Background	May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200