Application
Fig1: Western blot analysis of Smad3 on different lysates with Rabbit anti-Smad3 antibody at 1/20,000 dilution. Lane 1: HeLa cell lysate, Lane 2: A549 cell lysate, Lane 3: HL-60 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: bEnd.3 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute 52 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 1/20,000 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.Application
Fig2:Immunocytochemistry analysis of A549 cells labeling Smad3 with Rabbit anti-Smad3 antibody 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-Smad3 antibody at 1/500 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 1/1,000 dilution.Application
Fig3: ICC staining Smad3 in SW480 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig4: ICC staining Smad3 in HepG2 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig5: Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-Smad3 antibody. Counter stained with hematoxylin.Application
Fig6: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Smad3 antibody. Counter stained with hematoxylin.Application
Fig7: Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Smad3 antibody. Counter stained with hematoxylin.Application
Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.Application
Fig9: Flow cytometric analysis of HeLa cells labeling Smad3. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Smad3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Smad proteins, the mammalian homologs of the Drosophila Mothers against dpp (Mad) have been implicated as downstream effectors of TGFβ/BMP signaling. Smad1 (also designated Madr1 or JV4-1), Smad5 and mammalian Smad8 (also designated Smad9 or MADH6) are effectors of BMP2 and BMP4 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 family members. |
| Immunogen | Synthetic peptide within Human Smad3 aa 181-230 / 425. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:5000-1:20000 ICC/IF:1:500 IHC:1:50-1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | DKFZP586N0721 antibody
DKFZp686J10186 antibody
hMAD 3 antibody
hMAD-3 antibody
hSMAD3 antibody
HSPC193 antibody
HST17436 antibody
JV15 2 antibody
JV15-2 antibody
JV152 antibody
LDS1C antibody
LDS3 antibody
MAD (mothers against decapentaplegic Drosophila) homolog 3 antibody
MAD homolog 3 antibody
Mad homolog JV15 2 antibody
Mad protein homolog antibody
MAD mothers against decapentaplegic homolog 3 antibody Mad3 antibody MADH 3 antibody MADH3 antibody MGC60396 antibody Mothers against decapentaplegic homolog 3 antibody Mothers against DPP homolog 3 antibody SMA and MAD related protein 3 antibody SMAD 3 antibody SMAD antibody SMAD family member 3 antibody SMAD mothers against DPP homolog 3 antibody Smad3 antibody SMAD3_HUMAN antibody |
|---|---|
| Molecular Weight(MW) | 48kDa(Observed band size:55kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
Application
Fig1: Western blot analysis of Smad3 on different lysates with Rabbit anti-Smad3 antibody at 1/20,000 dilution. Lane 1: HeLa cell lysate, Lane 2: A549 cell lysate, Lane 3: HL-60 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: bEnd.3 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute 52 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 1/20,000 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.
Application
Fig2:Immunocytochemistry analysis of A549 cells labeling Smad3 with Rabbit anti-Smad3 antibody 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-Smad3 antibody at 1/500 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 1/1,000 dilution.
Application
Fig3: ICC staining Smad3 in SW480 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig4: ICC staining Smad3 in HepG2 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig5: Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-Smad3 antibody. Counter stained with hematoxylin.
Application
Fig6: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Smad3 antibody. Counter stained with hematoxylin.
Application
Fig7: Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Smad3 antibody. Counter stained with hematoxylin.
Application
Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes 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.
Application
Fig9: Flow cytometric analysis of HeLa cells labeling Smad3. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:5000-1:20000 ICC/IF:1:500 IHC:1:50-1:200 FC:1:1000 |
|---|
| 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. |
