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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\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.
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\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.
Bioss unconjugated antibody vial

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-100uL
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
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

Frequently asked questions

Which applications is phospho-Smad2 (Ser465) Polyclonal Antibody (bs-2224R) validated for, and at what dilution?

bs-2224R is validated for 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). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does phospho-Smad2 (Ser465) Polyclonal Antibody (bs-2224R) react with?

Tested reactivity: Human, Mouse. Predicted reactivity (expected from sequence homology but not yet tested): Rat, Dog, Cow, Horse, Chicken.

How should bs-2224R be stored?

Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise phospho-Smad2 (Ser465) Polyclonal Antibody (bs-2224R)?

KLH conjugated synthetic phosphopeptide derived from human Smad2 around Ser465

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