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Proteins and peptidesAnti-Ly6g antibody [1A8] - mouse IgG2c (Chimeric)
Low endotoxin, Azide free.
Our first-to-market chimera with mouse IgG2c backbone, this functional antibody specifically depletes neutrophils in vivo for up to 72h.
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Rabbit Recombinant Monoclonal SMAD4 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 170 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | IP | Flow Cyt | WB | ICC/IF | |
---|---|---|---|---|---|
Human | Tested | Not recommended | Not recommended | Tested | Not recommended |
Mouse | Expected | Not recommended | Not recommended | Tested | Not recommended |
Rat | Expected | Not recommended | Not recommended | Tested | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes For unpurified use at 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Rat | Dilution info - | Notes For unpurified use at 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Human | Dilution info - | Notes For unpurified use at 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/5000 | Notes - |
Species Rat | Dilution info 1/5000 | Notes - |
Species Human | Dilution info 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes For unpurified use at 1/100 - 1/250. |
Species Rat | Dilution info - | Notes For unpurified use at 1/100 - 1/250. |
Species Human | Dilution info - | Notes For unpurified use at 1/100 - 1/250. |
In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions (By similarity). Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling (PubMed:25514493). Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD family member 4, SMAD 4, Smad4, hSMAD4, MADH4, DPC4, SMAD4
Rabbit Recombinant Monoclonal SMAD4 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 170 publications.
Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD family member 4, SMAD 4, Smad4, hSMAD4, MADH4, DPC4, SMAD4
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EP618Y
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Lane 1: Wild type HAP1 whole cell lysate (20 μg)
Lane 2: SMAD4 knockout HAP1 whole cell lysate (20 μg)
Lane 3: HepG2 whole cell lysate (20 μg)
Lane 4: Jurkat whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759)
Predicted band size: 60 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution
Lane 1: SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg
Lane 2: Ramos (Human Burkitt's lymphoma cell line) cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution
Lane 1: NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg
Lane 2: Mouse embryo tissue lysate at 20 µg
Lane 3: Mouse skin tissue lysate at 20 µg
Lane 4: Mouse lung tissue lysate at 20 µg
Lane 5: PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 20 µg
Lane 6: C6 cell lysate at 20 µg
Lane 7: Rat skin tissue lysate at 20 µg
Lane 8: Rat lung tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Exposure time: 3min
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution
Lane 1: SW480 (Human colorectal adenocarcinoma cell line) cell lysate at 20 µg
Lane 2: HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg
Lane 4: Human skin tissue lysate at 20 µg
Lane 5: Human lung tissue lysate at 20 µg
Lane 6: Human artery tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Exposure time: 30s
Different batches of ab40759 were tested on Ramos (Human Burkitt's lymphoma B lymphocyte) lysate at 0.7 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 60 kDa.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759)
Predicted band size: 60 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/10000 dilution
All lanes: NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution
All lanes: PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution
All lanes: SHSY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg
Predicted band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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