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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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Knockout Tested Rabbit Recombinant Monoclonal SMAD2 phospho S465 + S467 antibody. Suitable for Dot, WB, IP, Flow Cyt (Intra), ICC/IF and reacts with Synthetic peptide - Human, Human, Mouse, Rat samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
Dot | WB | IP | Flow Cyt (Intra) | ICC/IF | IHC-P | ChIC/CUT&RUN | |
---|---|---|---|---|---|---|---|
Human | Expected | Tested | Tested | Tested | Tested | Not recommended | Not recommended |
Mouse | Expected | Tested | Tested | Tested | Tested | Not recommended | Not recommended |
Rat | Expected | Tested | Expected | Not recommended | Not recommended | Not recommended | Not recommended |
Synthetic peptide - Human | Tested | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species Mouse | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/5000 | Notes - |
Species Mouse | Dilution info 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species Mouse | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Synthetic peptide - Human | Dilution info - | Notes - |
Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma. 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 2, MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, SMAD family member 2, hMAD-2, SMAD 2, Smad2, hSMAD2, MADR2, MADH2, SMAD2
Knockout Tested Rabbit Recombinant Monoclonal SMAD2 phospho S465 + S467 antibody. Suitable for Dot, WB, IP, Flow Cyt (Intra), ICC/IF and reacts with Synthetic peptide - Human, Human, Mouse, Rat samples.
Mothers against decapentaplegic homolog 2, MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, SMAD family member 2, hMAD-2, SMAD 2, Smad2, hSMAD2, MADR2, MADH2, SMAD2
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR28610-44
Affinity purification Protein A
Not suitable for rat FC and ICC/IF.
When tested in dot blot this antibody recognizes Smad2 phosphorylated at both S465 and S467 and does not cross-react with Smad2 when singly phosphorylated at S465 or S467.
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.
Smad2 acts as an intracellular mediator for TGF-β signaling a pathway important for regulating cell proliferation differentiation and apoptosis. Smad2 typically functions as part of a heteromeric complex with Smad4 another key player in TGF-β signaling. Upon activation phosphorylated Smad2 combines with Smad4 to form a complex that translocates into the nucleus. This complex then binds to specific DNA sequences to regulate the transcription of target genes involved in processes such as cell growth inhibition and extracellular matrix production.
Smad2 also known as Mothers against decapentaplegic homolog 2 (MAD2) or MADR2 is a signaling protein involved in the transforming growth factor-beta (TGF-β) receptor pathway. Smad2 has a molecular mass of approximately 58 kDa and expresses in various tissues including epithelial mesenchymal and endothelial cells. Smad2 undergoes phosphorylation on serine residues in response to TGF-β signaling converting it into phosphorylated forms often referred to as p-Smad2 or phospho-Smad2. These phosphorylated forms are critical for the relay of signals from the cell surface to the nucleus.
Smad2 is integral to the TGF-β and activin receptor signaling pathways. These pathways are essential in controlling cell growth and immune responses. Smad2 interacts with other proteins such as Smad3 in addition to Smad4 to modulate gene expression effectively. The interaction between Smad2 and these proteins ensures precise control of cellular responses to external stimuli emphasizing its pivotal role in maintaining cellular homeostasis.
Smad2 correlates with various pathological conditions including fibrosis and cancer. Aberrant Smad2 signaling can contribute to the development of these diseases as excessive TGF-β signaling promotes fibrotic tissue deposition and tumor progression. Smad2 connects with other proteins like Smad3 in these pathological contexts both acting as mediators of abnormal cellular behaviors. Understanding the regulatory mechanisms of Smad2 can help develop therapeutic strategies against disorders linked to dysregulated TGF-β signaling.
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.
The molecular weight observed is consistent with what has been described in the literature (PMID: 34655614).
The up-regulation of pSmad2 is induced by TGF-beta1 treatment (PMID: 34655614, 24347165).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (AB316117) at 1/1000 dilution
Lane 1: Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 2: NIH/3T3 treated with 20 ng/ml TGF beta 1 for 15 minutes whole cell lysate at 20 µg
Lane 3: Untreated C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 4: C6 treated with 10 ng/ml TGF beta 3 for 30 minutes whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution
Observed band size: 60 kDa, 36 kDa
Exposure time: 70s
The molecular weight observed is consistent with what has been described in the literature (PMID: 34655614).
The up-regulation of pSmad2 is induced by TGF-beta treatment (PMID: 34655614, 24347165).
In Western blot, ab316117 was shown to bind specifically to Smad2. Target of interest was observed at 60 kDa in wild-type Hela cell lysates (lanes 2) with no signal observed at this size in Smad2 knockout cell line lysates (lanes 3-4 KO cell line ab255430/KO cell lysate ab263833 ).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Smad2 antibody [EP784Y] - Total protein control (ab40855) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (AB316117) at 1/1000 dilution
Lane 1: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: Wild-type Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (untreated membrane) at 20 µg
Lane 3: Untreated Smad2 knockout Hela whole cell lysate (untreated membrane) at 20 µg
Lane 4: Smad2 knockout Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (untreated membrane) at 20 µg
Lane 5: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 6: Wild-type Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 7: Untreated Smad2 knockout Hela whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 8: Smad2 knockout Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 60 kDa, 36 kDa
Exposure time: 37s
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labellingSmad2 (phospho S465 + S467) with ab316117 at 1/500 (1.014 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution.
Confocal image showing nuclear staining in NIH/3T3 cells (shown in green) treated with TGF-β1 (20 ng/mL) for 15 min. The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution. The Nuclear counterstain was DAPI.
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SMAD2 KO HeLa (human cervical adenocarcinoma epithelial cell) (ab255430) cells labellingSmad2 (phospho S465 + S467) with ab316117 at 1/500 (1.014 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution.
Confocal image showing nuclear and weak cytoplasmic staining in parental HeLa cells (shown in green) treated with TGF-β1 (20 ng/mL) for 1 hr. The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution. The Nuclear counterstain was DAPI.
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized (SMAD2 KO HeLa (human cervical adenocarcinoma epithelial cell) treated with 20ng/ml TGF-β1 for 1h (Red) and untreated SMAD2 KO HeLa (dotted red)) (Left) / (Parental HeLa treated with 20ng/ml TGF-β1 for 1h (Red) and untreated HeLa (dotted red)) (Right) cells labelling Smad2 (phospho S465 + S467) with ab316117 at 1/5000 dilution (0.01 ug)/Red and Dotted Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
Dot blot analysis of Smad2 (phospho S465 + S467) using ab316117 at 1:1000 (0.507 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:100,000 dilution.
The antibody recognizes Smad2 phosphorylated at S465 and S467. It does not cross-react with Smad2 when singly phosphorylated at S465 or S467.
All lanes: Dot Blot - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (AB316117) at 1/1000 dilution
Lane 1: Smad2 (phospho S465+S467) peptide
Lane 2: Smad2 (phospho S465) peptide
Lane 3: Smad2 (phospho S467) peptide
Lane 4: Smad2 non-phospho peptide
All lanes: Dot Blot at 1/100000 dilution
Exposure time: 180s
Smad2 (phospho S465 + S467) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) treated with 20ng/ml TGF beta1 for 15 minutes whole cell lysate with ab316117 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316117 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) treated with 20ng/ml TGF beta1 for 15 minutes, whole cell lysate
Lane 2: ab316117 IP in NIH/3T3 (mouse embryonic fibroblast) treated with 20ng/ml TGF beta1 for 15 minutes whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab316117 in NIH/3T3 treated with 20ng/ml TGF beta1 for 15 minutes whole cell lysate
All lanes: Immunoprecipitation - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (AB316117) at 1/30 dilution
All lanes: NIH/3T3 (mouse embryonic fibroblast) treated with 20ng/ml TGF beta1 for 15 minutes whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (AB131366) at 1/5000 dilution
Exposure time: 180s
Smad2 (phospho S465 + S467) was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) treated with 20ng/ml TGF beta1 for 60 minutes whole cell lysate with ab316117 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316117 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 20ng/ml TGF beta1 for 60 minutes, whole cell lysate
Lane 2: ab316117 IP in HeLa (human cervix adenocarcinoma epithelial cell) treated with 20ng/ml TGF beta1 for 60 minutes whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab316117 in HeLa treated with 20ng/ml TGF beta1 for 60 minutes whole cell lysate
All lanes: Immunoprecipitation - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (AB316117) at 1/30 dilution
All lanes: HeLa (human cervix adenocarcinoma epithelial cell) treated with 20ng/ml TGF beta1 for 60 minutes whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (AB131366) at 1/5000 dilution
Exposure time: 58s
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) treated with 20ng/ml TGF beta 1 for 15 min (Red) / untreated NIH/3T3 (dotted red) cells labelling Smad2 (phospho S465 + S467) with ab316117 at 1/5000 dilution (0.01 ug)/Red and Dotted Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
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