phospho-Smad2 (Ser465) Polyclonal Antibody

$Raji cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad2 (Ser465) Polyclonal Antibody(bs-2224R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Raji 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 (Ser465) Polyclonal Antibody(bs-2224R)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).

$Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103 and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with phospho-Smad2 (Ser465) Polyclonal Antibody(bs-2224R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Hela 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 phospho-Smad2 (Ser465) Polyclonal Antibody(bs-2224R)at 1:100 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).

Lane 1: Human Hela cell lysates; Lane 2: Human HT1080 cell lysates; Lane 3: Human Jurkat cell lysates; Lane 4: Human HL-60 cell lysates probed with Smad2 Polyclonal Antibody, Unconjugated (bs-2224R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Applications

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

Reactivity

Human
Mouse

Predicted Reactivity

Rat
Dog
Cow
Horse
Chicken

Overview
Catalog #	bs-2224R
Product Name	phospho-Smad2 (Ser465) Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)
Reactivity	Human, Mouse
Predicted Reactivity	Rat, Dog, Cow, Horse, Chicken
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human Smad2 around Ser465
Modification Site	Ser465
Clonality	Polyclonal
Clone #	#REF!
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	4087
Subcellular location	Cytoplasm, Nucleus
Synonyms	Smad2Ser465; p-Smad2Ser465; Smad2S465; Mothers against decapentaplegic homolog 2; SMAD 2; Mothers against DPP homolog 2; Smad2; hMAD 2; hSMAD2; JV18 1; JV18; JV181; MAD; MAD Related Protein 2; MADH2; MADR2; MGC22139; MGC34440; Mothers Against Decapentaplegic Homolog 2; mothers against DPP homolog 2; SMAD 2; SMAD; SMAD2; SMAD2_HUMAN.
Background	SMAD2 or Mothers against decapentaplegic homolog 2 is a polypeptide that, as its name describes, is a homolog of the Drosophila gene: "Mothers against decepentaplegic". It belongs to the SMAD family of proteins, which belong to the TGF-Beta superfamily of modulators. Like many other TGF-Beta family members SMAD2 is involved in cell signalling. SMAD2 modulates signals of activin and TGF-Beta's. It interacts with SMAD anchor for receptor activation (SARA). The binding of ligands causes the phosphorylation of the SMAD2 protein and the dissociation from SARA and the association with SMAD4. It is subsequently transferred to the nucleus where it forms complexes with other proteins and acts as a transcription factor. SMAD2 is a receptor regulated SMAD (R-SMAD) and is activated by bone morphogenetic protein type 1 receptor kinase. Smad2 (Mothers against decapentaplegic homolog 2; SMAD 2; Mothers against DPP homolog 2;)
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