WB result of Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: 293T whole cell lysate 20 µg
Lane 2: 293T whole cell lysate 20 µg (phosphatase treated membrane)
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 52 kDa
Observed MW: 52, 56 kDa
Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb (S-3372-136)
Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb (S-3372-136)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-SMAD2/3 (Thr8) |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | Q15796、P84022 |
| Clone Number | S-3372-136 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ChIP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | 293T, RAW264.7, PC-12 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt |
| ChIP | 1:20-1:50 | Hu |
Background
Phospho-SMAD2/3 (Thr8) refers to the phosphorylation of SMAD2 and SMAD3 proteins at threonine 8, a site distinct from their canonical C-terminal SXS motif phosphorylation by TGF-β receptors. This alternative phosphorylation is typically mediated by MAP kinases (ERK, JNK, p38) or CDKs in response to various stimuli including growth factors, stress, and cell cycle signals. It modulates SMAD transcriptional activity, stability, and subcellular localization, serving as a regulatory crossroad between TGF-β signaling and other pathways. Aberrant Thr8 phosphorylation has been implicated in cancer progression and fibrotic diseases, influencing cell proliferation, migration, and epithelial-mesenchymal transition.
Picture
Picture
Western Blot
WB result of Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: RAW264.7 whole cell lysate 20 µg
Lane 2: RAW264.7 whole cell lysate 20 µg (phosphatase treated membrane)
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 52 kDa
Observed MW: 56 kDa
WB result of Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Lane 2: PC-12 whole cell lysate 20 µg (phosphatase treated membrane)
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 52 kDa
Observed MW: 56 kDa
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HaCaT+Htgf-beta3(7ng/ml, 1h) (+) cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb (S-3372-136) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:20 for immunoprecipitation. Post- immunoprecipitation, both samples were washed, eluted, and cross-links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of ID1 and SAT-α in Phospho-SMAD2/3 (Thr8) Recombinant Rabbit mAb
(S-3372-136)-immunoprecipitated sample.
