Phospho-Smad2 (S250) Recombinant Rabbit Monoclonal Antibody [SD207-1]
cat.: ET1612-32
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SD207-1 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 52 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser250 of Human Smad2 aa 221-270 / 467. |
| Positive control: | HeLa cell lysate, HeLa treated with 200nM PMA for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate, C6 cell lysate, C6 treated with 200nM PMA for 30 minutes cell lysate, HeLa. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB FC IP |
1:5,000 1:50-1:100 1-2μg/sample |
| Uniprot #: | SwissProt: Q15796 Human | Q62432 Mouse | O70436 Rat |
| Alternative names: | Drosophila, homolog of, MADR2 hMAD-2 HsMAD2 JV18 JV18-1 JV181 MAD MAD homolog 2 MAD Related Protein 2 Mad-related protein 2 MADH2 MADR2 MGC22139 MGC34440 Mother against DPP homolog 2 Mothers against decapentaplegic homolog 2 Mothers against decapentaplegic, Drosophila, homolog of, 2 Mothers against DPP homolog 2 OTTHUMP00000163489 Sma and Mad related protein 2 Sma- and Mad-related protein 2 MAD SMAD 2 SMAD family member 2 SMAD, mothers against DPP homolog 2 SMAD2 SMAD2_HUMAN |
Images
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Fig1:
Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody (ET1612-32) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 200nM PMA for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 55 kDa Exposure time: 30 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-32) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody (ET1612-32) at 1/5,000 dilution. Lane 1: C6 cell lysate Lane 2: C6 treated with 200nM PMA for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52 kDa Observed band size: 55 kDa Exposure time: 2 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-32) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Phospho-Smad2 (S250) was immunoprecipitated from 0.2 mg HeLa treated with 100nM TPA for 30 minutes cell lysate with ET1612-32 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1612-32 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 100nM TPA for 30 minutes cell lysate (input) Lane 2: ET1612-32 IP in HeLa treated with 100nM TPA for 30 minutes cell lysate Lane 3: Rabbit IgG instead of ET1612-32 in HeLa treated with 100nM TPA for 30 minutes cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 35 seconds; ECL: K1802 |
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Fig4:
Flow cytometric analysis of HeLa cells labeling Phospho-Smad2 (S250). Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-32, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.