WB
Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody 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. Exposure time: 30 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.FC
Flow cytometric analysis of Hela cells with Phospho-Smad2 (S250) antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibodyIHC
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.IP
Phospho-Smad2 (S250) was immunoprecipitated from 0.2 mg HeLa treated with 100nM TPA for 30 minutes cell lysate with Rabbit anti-Phospho-Smad2 (S250) antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Phospho-Smad2 (S250) antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody 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: Rabbit anti-Phospho-Smad2 (S250) antibody IP in HeLa treated with 100nM TPA for 30 minutes cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Phospho-Smad2 (S250) antibody in HeLa treated with 100nM TPA for 30 minutes cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 35 seconds.| Product Name | Phospho-Smad2 (S250) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Smad proteins, the mammalian homologs of the Drosophila mothers against decapentaplegic (Mad), have been implicated as downstream effectors of TGFβ/BMP signaling. Smad1 (also designated Madr1 or JV4-1) and Smad5 are effectors of BMP-2 and BMP-4 function, while Smad2 (also designated Madr2 or JV18-1) and Smad3 are involved in TGFβ and Activin-mediated growth modulation. Smad4 (also designated DPC4) has been shown to mediate all of the above activities through interaction with various Smad family members. Smad6 and Smad7 regulate the response to Activin/TGFβ signaling by interfering with TGFβ-mediated phosphorylation of other Smad proteins. |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser250 of Human Smad2 aa 221-270 / 467. |
| Modification | p-S250 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCIHCWB |
| WB:1:5000 FC:1:50-1:100 IHC:1:50 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Drosophila homolog of MADR2 antibody hMAD-2 antibody HsMAD2 antibody JV18 antibody JV18-1 antibody JV181 antibody MAD antibody MAD homolog 2 antibody MAD Related Protein 2 antibody Mad-related protein 2 antibody MADH2 antibody MADR2 antibody MGC22139 antibody MGC34440 antibody Mother against DPP homolog 2 antibody Mothers against decapentaplegic homolog 2 antibody Mothers against decapentaplegic Drosophila homolog of 2 antibody Mothers against DPP homolog 2 antibody OTTHUMP00000163489 antibody Sma and Mad related protein 2 antibody Sma- and Mad-related protein 2 MAD antibody SMAD 2 antibody SMAD family member 2 antibody SMAD mothers against DPP homolog 2 antibody SMAD2 antibody SMAD2_HUMAN antibody |
|---|---|
| Molecular Weight(MW) | 52kDa(Observed band size: 55kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of Phospho-Smad2 (S250) on different lysates with Rabbit anti-Phospho-Smad2 (S250) antibody 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. Exposure time: 30 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
FC
Flow cytometric analysis of Hela cells with Phospho-Smad2 (S250) antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody
IHC
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Phospho-Smad2 (S250) antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/50 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.
IP
Phospho-Smad2 (S250) was immunoprecipitated from 0.2 mg HeLa treated with 100nM TPA for 30 minutes cell lysate with Rabbit anti-Phospho-Smad2 (S250) antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Phospho-Smad2 (S250) antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody 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: Rabbit anti-Phospho-Smad2 (S250) antibody IP in HeLa treated with 100nM TPA for 30 minutes cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Phospho-Smad2 (S250) antibody in HeLa treated with 100nM TPA for 30 minutes cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 35 seconds.| Application Notes | WB:1:5000 FC:1:50-1:100 IHC:1:50 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
