Product nameAnti-SMAD1/3/5 (phospho S423 + S425) antibody
DescriptionRabbit polyclonal to SMAD1/3/5 (phospho S423 + S425)
Specificityab51451 is specific for the phosphorylated form of the protein although minimal reactivity with nonphosphorylated human Smad3 can be observed by ELISA and WB. ab51451 also reacts with phosphorylated Smad1 and Smad5 but negligible reactivity is seen against other phosphorylated Smad family members.
Tested applicationsSuitable for: IHC-P, WB, ELISA, ICC/IFmore details
Species reactivityReacts with: Mouse, Chicken, Human
Predicted to work with: Rat, Cow, Pig, Xenopus laevis, Zebrafish
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.33% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
Purification notesab51451 was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.
Our Abpromise guarantee covers the use of ab51451 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. PubMed: 20519446|
|WB||1/2000 - 1/20000. Predicted molecular weight: 48 kDa. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.|
|ELISA||1/25000 - 1/100000.|
|ICC/IF||Use at an assay dependent concentration.|
- BSP1 antibody
- MADH1 antibody
- MADH3 antibody
All lanes : Anti-SMAD1/3/5 (phospho S423 + S425) antibody (ab51451) at 1 µg/ml
Lane 1 : HEK293 lysate
Lane 2 : HeLa Lysate
Lane 3 : MCF-7 Lysate
Lane 4 : Jurkat Lysate
Lane 5 : A431 Lysate
Lane 6 : A549 Lysate
Lane 7 : LNCap Lysate
Lane 8 : MOLT-4 Lysate
Lane 9 : Ramos Lysate
Lane 10 : Raji Lysate
Lane 11 : A-172 Lysate
Lane 12 : NIH/3T3 Lysate
Lysates/proteins at 10 µg per lane.
Lanes 1-2 & 4-12 : Peroxidase rabbit secondary antibody at 1/30000 dilution
Lane 3 : Peroxidase rabbit secondary antibody
Predicted band size: 48 kDa
ab51451 at 2.5 µg/ml staining epidermal keratinocytes at 40X. This image shows strong nuclear staining with localization of the antibody as the precipated red siganl and a hematoxylin purple nuclear counterstain.
ab51451 staining Smad3 (phospho S423 + S425) in PC3 and LnCAP cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 70% ethanol and blocked for 1 hour at 26°C. Samples were incubated with primary antibody (1/150) for 16 hours at 4°C. An IRDye® 800CW-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. NC-PC3 - negative control PC3 cells, NC-LnCAP - negative control LnCAP cells.
All lanes : Anti-SMAD1/3/5 (phospho S423 + S425) antibody (ab51451) at 1/2000 dilution
Lane 1 : MEF lysate (control)
Lane 2 : MEF lysate, after stimulation with 2 ng/ml of TGF-ß for 1 hour
Lane 3 : MEF-/-Smad3 lysate (control)
Lane 4 : MEF-/-Smad3 lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
Lane 5 : Hep3B lysate (control)
Lane 6 : Hep3B lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
Lane 7 : HEK 293 lysate (control)
Predicted band size: 48 kDa
Image courtesy of Ying Zhang, NIH, CCR, Bethesda, MD.
This product has been referenced in:
- Murugavel S et al. Valproic Acid Induces Endothelial-to-Mesenchymal Transition-Like Phenotypic Switching. Front Pharmacol 9:737 (2018). Read more (PubMed: 30050438) »
- Boswell BA et al. Dual function of TGFß in lens epithelial cell fate: implications for secondary cataract. Mol Biol Cell 28:907-921 (2017). Read more (PubMed: 28209733) »