WB
Western blot analysis using Smad3 antibody against Hela(1),A549(2),Mcf-7(3),Mouse liver tissue(4),Mouse lung tissue(5),Mouse spleen tissue(6) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP（OM644366）, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60SIF-P
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:150 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:150 and incubate for one hour. Tyramine labeled with 488 (Green,OM642679) was used as chromogenic agent, and DAPI(Blue,OM642679) was used for double dyeing, and the anti-fluorescence attenuation tablets were sealed.IHC
Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded lung cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded ovarian cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded prostate cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ICC/IF
Immunofluorescence analysis of C2C12 cells using Smad3 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutesELISA
Blue line: Antibody; Orange line: Isotype control antibody| Product Name | Anti-Smad3 antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Mouse |
| Tested Applications | ELISAICC/IFIF-PIHCWB |
| WB:1:1000-1:2000 IF-P:1:200-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | SMAD3 |
|---|---|
| Gene Synonyms | 7120426M23Rik CHTD8 hMAD-2 hMAD-3 hSMAD2 hSMAD3 HSPC193 HsT17436 JV15-2 JV18 JV18-1 LDS1C LDS3 LDS6 mad3 MADH2 MADH3 MADR2 mMad2 Smad 3 Smad-2 |
| Gene Full Name | SMAD family member 3 |
| Gene Summary | The SMAD family of proteins are a group of intracellular signal transducer proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. The SMAD3 protein functions in the transforming growth factor-beta signaling pathway, and transmits signals from the cell surface to the nucleus, regulating gene activity and cell proliferation. This protein forms a complex with other SMAD proteins and binds DNA, functioning both as a transcription factor and tumor suppressor. Mutations in this gene are associated with aneurysms-osteoarthritis syndrome and Loeys-Dietz Syndrome 3. [provided by RefSeq, May 2022] |
| Alternative Names | DKFZP586N0721 MAD (mothers against decapentaplegic Drosophila) homolog 3 MAD homolog 2 MAD homolog 3 mad homolog JV15-2 mad protein homolog MAD mothers against decapentaplegic homolog 3 mad-related protein 2 mad3 mMad3 mother against DPP homolog 2 mothers against decapentaplegic homolog 2 mothers against decapentaplegic homolog 3 mothers against DPP homolog 2 mothers against DPP homolog 3 pSMAD2 Sma- and Mad-related protein 2 SMA- and MAD-related protein 3 SMAD SMAD 2 Smad 3 SMAD mothers against DPP homolog 2 SMAD mothers against DPP homolog 3 |
| Molecular Weight(MW) | 48kDa |
| Cellular Localization | Cytoplasm,Nucleus. |
| SwissProt ID | P84022 |
|---|
WB
Western blot analysis using Smad3 antibody against Hela(1),A549(2),Mcf-7(3),Mouse liver tissue(4),Mouse lung tissue(5),Mouse spleen tissue(6) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP（OM644366）, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S
IF-P
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:150 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:150 and incubate for one hour. Tyramine labeled with 488 (Green,OM642679) was used as chromogenic agent, and DAPI(Blue,OM642679) was used for double dyeing, and the anti-fluorescence attenuation tablets were sealed.
IHC
Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded lung cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded ovarian cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded prostate cancer tissues using Smad3 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ICC/IF
Immunofluorescence analysis of C2C12 cells using Smad3 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes
ELISA
Blue line: Antibody; Orange line: Isotype control antibody| Application Notes | WB:1:1000-1:2000 IF-P:1:200-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | PBS ONLY |
