Smad4 (3A1) Monoclonal Antibody

Lane 1: PC-12 lysates probed with Smad4 (3A1) Monoclonal Antibody (bsm-52225R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.

Hela cells were stained with Smad4 (3A1) Monoclonal Antibody (bsm-52225R) at [1:200] incubated overnight at 4C followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

$Paraformaldehyde-fixed, paraffin embedded mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with Smad4 (3A1) Monoclonal Antibody, (bsm-52225R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded mouse brain; 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 Smad4 (3A1) Monoclonal Antibody, (bsm-52225R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

Lane 1: Lung lysates probed with Smad4 (3A1) Monoclonal Antibody, Unconjugated (bsm-52225R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Lane 1: Hela cell lysates; Lane 2: Jurkat cell lysates; Lane 3: HepG2 cell lysates; Lane 4: HL60 cell lysates ; Lane 5:Mouse Cerebrum lysates probed with Smad4 Monoclonal Antibody, Unconjugated (bsm-52225R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Lane 1: Human Hela cell lysates; Lane 2: Human Jurkat cell lysates; Lane 3:Human HepG2 cell lysates; Lane 4: Human HL-60 cell lysates; Lane 5: Mouse Cerebrum lysates probed with Smad4 monoclonal Antibody, Unconjugated (bsm-52225R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

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).

Applications

WB
IHC-P
IF(ICC)

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52225R
Product Name	Smad4 (3A1) Monoclonal Antibody
Applications	WB, IHC-P, IF(ICC)
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant human Smad4 protein, around N-terminal 1-200aa.
Clonality	Monoclonal
Clone #	3A1
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	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
IF(ICC)	1:50-200