WB
Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody at 1/5000 dilution. Lane 1: HeLa cell lysate, Lane 2: HT-29 cell lysate, Lane 3: Jurkat cell lysate, Lane 4: HL-60 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute 20 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 1/5000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody at 1/1000 dilution. Lane 1: Jurkat cell lysate (15 µg/Lane), Lane 2: C2C12 cell lysate (15 µg/Lane), Lane 3: Mouse lung tissue lysate (30 µg/Lane), Lane 4: Mouse placenta tissue lysate (30 µg/Lane), Exposure time: 1 minute 21 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 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of Smad2 with anti-Smad2 antibody at 1:500 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad2 knockout HaCaT whole cell lysate (15 µg). Smad2 with anti-Smad2 antibody was shown to specifically react with Smad2 in wild-type HaCaT cells. NO band was observed when Smad2 knockout sample was tested. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (1:500) was used in 5% BSA at room temperature for 2 hours. Goat anti-Rabbit IgG-HRP antibody at 1:10,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HepG2 cells labeling Smad2 with Rabbit anti-Smad2 antibody at 1/100 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-Smad2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Smad2 with Rabbit anti-Smad2 antibody at 1/100 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-Smad2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-Smad2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Smad2 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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 cerebellum tissue with Rabbit anti-Smad2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 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.FC
Flow cytometric analysis of HepG2 cells labeling Smad2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (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 at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).FC
Flow cytometric analysis of NIH/3T3 cells labeling Smad2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (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 at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Smad2 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human Smad2 aa 220-270. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000-1:5000 ICC/IF:1:100 IHC:1:200-1:1000 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | SMAD2 |
|---|---|
| Gene Synonyms | JV18 LDS6 CHTD8 MADH2 MADR2 JV18-1 hMAD-2 hSMAD2 |
| Gene Full Name | SMAD family member 2 |
| Gene Summary | The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. This protein is recruited to the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation (SARA) protein. In response to TGF-beta signal, this protein is phosphorylated by the TGF-beta receptors. The phosphorylation induces the dissociation of this protein with SARA and the association with the family member SMAD4. The association with SMAD4 is important for the translocation of this protein into the nucleus, where it binds to target promoters and forms a transcription repressor complex with other cofactors. This protein can also be phosphorylated by activin type 1 receptor kinase, and mediates the signal from the activin. Alternatively spliced transcript variants have been observed for this gene. [provided by RefSeq, May 2012] |
| 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 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:58kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody at 1/5000 dilution. Lane 1: HeLa cell lysate, Lane 2: HT-29 cell lysate, Lane 3: Jurkat cell lysate, Lane 4: HL-60 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute 20 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 1/5000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody at 1/1000 dilution. Lane 1: Jurkat cell lysate (15 µg/Lane), Lane 2: C2C12 cell lysate (15 µg/Lane), Lane 3: Mouse lung tissue lysate (30 µg/Lane), Lane 4: Mouse placenta tissue lysate (30 µg/Lane), Exposure time: 1 minute 21 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 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Smad2 with anti-Smad2 antibody at 1:500 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad2 knockout HaCaT whole cell lysate (15 µg). Smad2 with anti-Smad2 antibody was shown to specifically react with Smad2 in wild-type HaCaT cells. NO band was observed when Smad2 knockout sample was tested. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (1:500) was used in 5% BSA at room temperature for 2 hours. Goat anti-Rabbit IgG-HRP antibody at 1:10,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HepG2 cells labeling Smad2 with Rabbit anti-Smad2 antibody at 1/100 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-Smad2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Smad2 with Rabbit anti-Smad2 antibody at 1/100 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-Smad2 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-Smad2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Smad2 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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 cerebellum tissue with Rabbit anti-Smad2 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 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.
FC
Flow cytometric analysis of HepG2 cells labeling Smad2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (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 at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
FC
Flow cytometric analysis of NIH/3T3 cells labeling Smad2. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (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 at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:5000 ICC/IF:1:100 IHC:1:200-1:1000 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
