antibody

Anti-Smad3 Antibody [SY25-01]

ET1607-41TR

20 uL

99 USD

monoclonal

domestic rabbit

nonconjugated

SY25-01

human, mouse, rat

western blot, flow cytometry, immunohistochemistry - paraffin section

Anti-Smad3 Antibody [SY25-01]

Human,Mouse,Rat

WB,IF-Cell,IF-Tissue,IHC-P,FC

Non-conjugated

Synthetic peptide within Human Smad3 aa 181-230 / 425.

SwissProt: P84022 Human;SwissProt: Q8BUN5 Mouse;SwissProt: P84025 Rat

Rabbit

SY25-01

IgG

20 uL

99 USD

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Liquid

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Protein A affinity purified.

Recombinant Rabbit monoclonal Antibody

HeLa cell lysate, A549 cell lysate, HL-60 cell lysate, NIH/3T3 cell lysate, bEnd.3 cell lysate, C6 cell lysate, Wild-type HaCaT whole cell lysate, Hela, SW480, HepG2, human lung carcinoma tissue, human breast carcinoma tissue, mouse liver tissue, mouse brain tissue, rat testis tissue.

Predicted band size: 48 kDa

Cytoplasm, Nucleus.

1 mg/mL.

WB: 1:5,000-1:20,000 ;IF-Cell: 1:50-1:200 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:50-1:200 ;FC: 1:50-1:100

ET1607-41_2.jpg

All lanes: Western blot analysis of Smad3 with anti-Smad3 antibody (ET1607-41) at 1:1,000 dilution. Lane 1: Wild-type HaCaT whole cell lysate (15 µg). Lane 2: Smad3 knockout HaCaT whole cell lysate (15 µg). ET1607-41 was shown to specifically react with Smad3 in wild-type HaCaT cells. NO band was observed when Smad3 knockout sample was tested. Wild-type and Smad3 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 (ET1607-41, 1:1,000) 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.

ET1607-41_3.jpg

ICC staining of Smad3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-41, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1607-41_4.jpg

ICC staining of Smad3 in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-41, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1607-41_5.jpg

ICC staining of Smad3 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-41, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1607-41_6.jpg

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1607-41, 1/50) 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.

ET1607-41_7.jpg

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1607-41, 1/50) 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.

ET1607-41_8.jpg

Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1607-41, 1/50) 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.

ET1607-41_9.jpg

Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Smad3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1607-41, 1/50) 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.

ET1607-41_10.jpg

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 (ET1607-41, 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.

ET1607-41_11.jpg

Flow cytometric analysis of Smad3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1607-41, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

ET1607-41_12.jpg

Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Smad3 antibody (ET1607-41) at 1/500 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 (ET1607-41) at 1/500 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.