SMAD2 Recombinant antibody

SMAD2 Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, rat

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240950A11

Cat no : 83841-5-RR

Synonyms

JV18 1, JV18, hSMAD2, hMAD-2, hMAD 2

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Validation Data Gallery

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  WB analysis using 83841-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83841-5-RR (SMAD2 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 83841-5-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using SMAD2 antibody (83841-5-RR, Clone: 240950A11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HeLa using 83841-5-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug SMAD2 Recombinant antibody (83841-5-RR, Clone:240950A11) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  FC experiment of Jurkat using 83841-5-RR

  1x10^6 Jurkat cells were intracellularly stained with 0.25 ug SMAD2 Recombinant antibody (83841-5-RR, Clone:240950A11) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83841-5-RR

  Biolayer interferometry (BLl) kinetic assays of 83841-5-RR against Human SMAD2 were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: HeLa cells, A549 cells, C6 cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: Jurkat cells, HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 83841-5-RR is SMAD2 Fusion Protein expressed in E. coli.

• Tested Reactivity: human, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: SMAD2 fusion protein Ag3237
• Full Name: SMAD family member 2
• Calculated Molecular Weight: 467 aa, 52 kDa
• Observed Molecular Weight: 58 kDa
• GenBank Accession Number: BC014840
• Gene Symbol: SMAD2
• Gene ID (NCBI): 4087
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

SMAD2, also named as MADH2 and MADR2, belongs to the dwarfin/SMAD family, contains 1 MH1 (MAD homology 1) domain and 1 MH2 (MAD homology 2) domain. SMAD2 is a receptor-regulated SMAD(R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta and activin type 1 receptor kinases. This protein may act as a tumor suppressor in colorectal carcinoma. It is phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, It is phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases, and then able to interact with SMURF2, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, it is phosphorylated on Ser-240 by CaMK2. It is phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. In response to TGF-beta, it is ubiquitinated by NEDD4L, which promotes its degradation. In response to TGF-beta signaling, it is acetylated on Lys-19 by coactivators, which increases transcriptional activity. The molecular weight of unphosphorylated forms of Smad2 is 52 kDa and phosphorylated forms of Smad2 is 58 kDa. (PMID: 9006934)