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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 antibody (ab28379)
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Western blot - Smad3 antibody (ab28379)
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ChIP - Smad3 antibody - ChIP Grade (ab28379)
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Product Name
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Smad3 antibody - ChIP Grade
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See all Smad3 antibodies (17)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Smad3 - ChIP Grade
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Immunogen
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Synthetic peptide, corresponding to amino acids 192-211 of Human Smad3
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Reacts with
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Hu, Ms
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Predicted to react
(species key)
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Rat
(Due to sequence homology)
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Specificity
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This antibody does not cross-react with other Smad proteins.
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Tested applications
(see key)
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ChIP, ICC/IF, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ChIP: Use at an assay dependent concentration.
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: 1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB: Use at an assay dependent dilution. Predicted molecular weight: 48 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Normal breast or breast carcinoma.
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Cellular localization
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In the cytoplasm in the absence of ligand; migration to the nucleus when complexed with SMAD4.
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Research areas
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Cancer >> Cancer Metabolism >> Response to hypoxia
Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Cancer >> Signal transduction >> Nuclear signaling >> SMAD family
Stem Cells >> Signaling Pathways >> TGF beta >> Cytoplasmic
Chromatin and Nuclear Signaling >> Nuclear Signaling Pathways >> SMADs
Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs
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Relevance
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Smad3 is a 50 kDa member of a family of proteins that act as key mediators of TGF beta superfamily signaling in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor regulated Smads, activin/TGF beta receptor regulated (Smad2 and 3) or BMP receptor regulated (Smad 1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smads; and the inhibitory Smads, (Smad6 and 7). Activated Smad3 oligomerizes with Smad4 upon TGF beta stimulation and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. Phosphorylation of the two TGF beta dependent serines 423 and 425 in the C terminus of Smad3 is critical for Smad3 transcriptional activity and TGF beta signaling.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.1% Sodium Azide
Constituents: 1% BSA, 10mM PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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This antibody is affinity purified.
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Form
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Liquid
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Concentration
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0.200 mg/ml
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Storage instructions
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Store at +4°C.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Smad3 antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Smad3 antibody - ChIP Grade images, references, products related to ab28379 and other tools.
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Smad3 antibody - ChIP Grade images:
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 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 antibody (ab28379)
Ab28379, at a 1/100 dilution, staining Smad3 in formalin fixed paraffin embedded human breast carcinoma sections by
Immunohistochemistry.
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Western blot - Smad3 antibody (ab28379)
All lanes : Smad3 antibody - ChIP Grade (ab28379) at 1/3000 dilution
Lane 1 : HGL-5 cells: no additives
Lane 2 : HGL-5 cells: + activin A
Lane 3 : HGL-5 cells: + Cyclophilin, siRNA Control
Lane 4 : HGL-5 cells: + activin A + Cyclophilin
Lane 5 : HGL-5 cells: non targeting, negativ control for siRNA
Lane 6 : HGL-5 cells: plus activin A, negative control for siRNA
Lane 7 : HGL-5 cells: plus siRNA Smartpool
Lane 8 : HGL-5 cells: + activin A +siRNA Smartpool
Lane 9 : Jurkat cells
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) (ab6721) at 1/50000 dilution
Predicted band size : 48 kDa
Observed band size : 58 kDa (why is the actual band size different from the predicted?)
This image is courtesy of an Abreview submitted by Mrs Birte Jablonski
See Abreview
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ChIP - Smad3 antibody - ChIP Grade (ab28379)
Chromatin was prepared from the nuclear cell lysate of a human myofibroblast cell line according to the X-ChIP protocol; cross-linking with formaldehyde for 10 minutes. ab28379 was diluted 1/16 (RIPA 0.1% SDS) and incubated with the sample for 16 hours at 4°C. ab28379 IP was used as the positive control and rabbit IgG pool IP was used as the negative control. Smad3 expression was enhanced by 2ng/ml TGFbeta 2. The immunoprecipitated DNA was quantified by real time PCR. This image is courtesy of an anonymous Abreview
See Abreview
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Immunocytochemistry/ Immunofluorescence-Smad3 antibody - ChIP Grade(ab28379)
ICC/IF image of ab28379 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28379, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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References for Smad3 antibody - ChIP Grade (ab28379)
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This product has been used in: (two most recent references)
Fu Y et al. Differential regulation of transforming growth factor {beta} signaling pathways by notch in human endothelial cells. J Biol Chem 284:19452-62 (2009). ChIP, IF; Mouse. PubMed: 19473993
Luo DD et al. Interleukin-1 beta regulates proximal tubular cell transforming growth factor beta-1 signalling. Nephrol Dial Transplant 24:2655-65 (2009). PubMed: 19420104
See all 10 publication references for this product.
If you publish research using ab28379 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Smad3
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Smad3 antibody - ChIP Grade - more information
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