Smad4 Recombinant Antibody

HUVEC cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad4 (3A1) Monoclonal Antibody(bsm-52205R)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).

$Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad4 Monoclonal Antibody, Unconjugated(bsm-52225R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$25 ug total protein per lane of various lysates (see on figure) probed with Smad4 monoclonal antibody, unconjugated (bsm-52225R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with Smad4 monoclonal antibody, unconjugated (bsm-52225R) 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

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52225r-TR
Product Name	Smad4 Recombinant Antibody
Applications	WB IHC-P IHC-F IF
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human Smad4 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	4089
Swiss Prot	Q13485
Synonyms	JIP; DPC4; MADH4; MYHRS; Mothers against decapentaplegic homolog 4; MAD homolog 4; Mothers against DPP homolog 4; Deletion target in pancreatic carcinoma 4; SMAD family member 4; SMAD 4; Smad4; hSMAD4
Background	In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions (By similarity). Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling. Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for syngernistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. 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