Smad2 Recombinant Antibody

$A431 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad2 (9A3) Monoclonal Antibody(bsm-52223R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

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 Smad2 (9A3) Monoclonal Antibody(bsm-52223R)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).

$Paraformaldehyde-fixed, paraffin embedded Rat kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat kidney; 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 Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.$

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\u00b0C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; 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 Smad2 (9A3) Monoclonal Antibody, Unconjugated (bsm-52223R) at 1:200 overnight at 4\u00b0C, DAB staining.

$The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-Smad2 antibody (bsm-52223R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-Smad2 antibody (bsm-52223R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated(bsm-52223R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$25 ug total protein per lane of various lysates (see on figure) probed with Smad2 monoclonal antibody, unconjugated (bsm-52223R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with Smad2 monoclonal antibody, unconjugated (bsm-52223R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$25 ug total protein per lane of various lysates (see on figure) probed with SMAD2 monoclonal antibody, unconjugated (bsm-52223R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with SMAD2 monoclonal antibody, unconjugated (bsm-52223R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt
ICC/IF

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52223r-TR
Product Name	Smad2 Recombinant Antibody
Applications	WB, IHC-P, IHC-F, IF, Flow-Cyt, ICC/IF
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human Smad2 protein, around N-terminal 1-200aa.
Clonality	Recombinant
Isotype	IgG
Concentration	Lot dependent
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	4087
Swiss Prot	Q15796
Synonyms	JV18; MADH2; MADR2; JV18-1; hMAD-2; hSMAD2; Mothers against decapentaplegic homolog 2; MAD homolog 2; Mothers against DPP homolog 2; Mad-related protein 2; SMAD family member 2; SMAD 2; Smad2
Background	Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC-F	1:100-500
IF
Flow-Cyt	1ug/Test
ICC/IF	1:50-100