STAT3(S727) (4G1) Monoclonal Antibody

$Lane 1: HeLa lysates; Lane 2: NIH\/3T3 lysates; probed with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.$

Lane 1: HeLa lysates; Lane 2: NIH\/3T3 lysates; probed with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.

$HeLa cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.$

HeLa cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$A431 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.$

A431 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$NIH\/3T3 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.$

NIH\/3T3 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with STAT3(S727) (4G1 ) Monoclonal Antibody (bsm-52210R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$Paraformaldehyde-fixed, paraffin embedded rat liver; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody (bsm-52210R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded rat liver; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody (bsm-52210R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

$Paraformaldehyde-fixed, paraffin embedded rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody (bsm-52210R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody (bsm-52210R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

$A431 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with STAT3(S727) (4G1) Monoclonal Antibody(bsm-52210R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

A431 cells(black) 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 STAT3(S727) (4G1) Monoclonal Antibody(bsm-52210R)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).

Lane 1:Rat Heart lysates; Lane 2:Rat Cerebrum lysates probed with phospho-STAT3 (Ser727) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat 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\u00b0C for 30min; Antibody incubation with STAT3(S727) (4G1) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:200 overnight at 4\u00b0C, DAB staining.

Lane 1: Human Hela cell lysates; Lane 2: Human A549 cell lysates probed with STAT3(S727) (4G1) Monoclonal Antibody, Unconjugated (bsm-52210R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-STAT3 (Ser727)) monoclonal Antibody, Unconjugated (bs-52210R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

A431 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-STAT3 (Ser727)) monoclonal Antibody, Unconjugated (bs-52210R) 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.

Applications

WB
FCM
IHC-P
IF(ICC)

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52210R
Product Name	STAT3(S727) (4G1) Monoclonal Antibody
Applications	WB, FCM, IHC-P, IF(ICC)
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Synthetic peptide derived from human STAT3(S727), around 700-750aa (phospho S727).
Modification Site	S727
Clonality	Monoclonal
Clone #	4G1
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	Aqueous buffered solution containing 1xTBS (pH7.4), 1%BSA, 40%Glycerol and 0.05% Sodium Azide.
Storage Condition	Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	6774
Swiss Prot	P40763
Synonyms	APRF; HIES; ADMIO; Signal transducer and activator of transcription 3; Acute-phase response factor; STAT3
Background	Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF and other growth factors. May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. Plays an important role in host defense in methicillin-resistant S.aureus lung infection by regulating the expression of the antimicrobial lectin REG3G (By similarity).
Application Dilution
WB	1:300-5000
FCM	1:20-100
IHC-P	1:200-400
IF(ICC)	1:50-200