Phospho-SMAD2 (S465+467) Recombinant Rabbit Monoclonal Antibody [PSH05-76]
cat.: HA722443
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | PSH05-76 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 52/48 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser465 and 467 of Human SMAD2. |
| Positive control: | HeLa cell lysate, HeLa starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate, NIH/3T3 starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate, HeLa cells treated with 20ng/mL TGF-β1 for 15 minutes, HeLa, NIH/3T3, human stomach tissue, rat lung tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:1,000 1:500 1:200 |
| 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 (S465+467) on different lysates with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52/48 kDa Observed band size: 60 kDa Exposure time: 1 minute 20 seconds; ECL: K1802; 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 (HA722443) at 1/1,000 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:
Immunocytochemistry analysis of HeLa cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunocytochemistry analysis of NIH/3T3 cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722443) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA722443) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722443) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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