Anti-pSmad1 Ser463/465 Antibody, clone 1N12 ZooMAb^® Rabbit Monoclonal

Quality Control Testing

Evaluated by Western Blotting in Jurkat cells treated with BMP2.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected Smad1 phosphorylated on serine 463/465 in lysate from serum starved Jurkat cells treated with BMP2 (50 ng/mL; 30 min.).

Tested applications

Peptide Inhibition Assay: Target band detection in Lysate from HeLa cells treated with BMP2 was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Affinity Binding Assay: A representative lot of this antibody bound phospho-Smad1-Ser463/465 with a KD of 2.4 x 10-7 in an affinity binding assay.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Clone 1N12 is a ZooMAb^® Rabbit recombinant monoclonal antibody that specifically detects Smad1 phosphorylated on serine 463/465.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Mothers against decapentaplegic homolog 1 (UniProt: Q15797; also known as MAD homolog 1, Mothers against DPP homolog 1, JV4-1, Mad-related protein 1, SMAD family member 1, SMAD1, Smad1, hSMAD1, Transforming growth factor-beta-signaling protein 1, BSP-1) is encoded by the SMAD1 (also known as BSP1, MADH1, MADR1) gene (Gene ID: 4086) in human. Smad1 is a transcriptional modulator that is activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. Smad1 is a receptor-regulated Smad (R-Smad) that is found in a complex with Smad4 and YY1. Upon phosphorylation at the C-terminal SVS motif (Ser463/465) by BMP type 1 receptor kinase it forms a trimer with another Smad1 and Smd4. The C-terminal phosphoserines of Smad1 are recognized by the Mad Homology 2 (MH2) domain of Smad4. The Smad1/Smad4 complex then translocates to the nucleus where Smad proteins bind to their cognate DNA binding sites with low affinity. This binding is further enhanced in the presence of transcriptional co-activators. It is exported from the nucleus to the cytoplasm upon dephosphorylation. Smad1 is ubiquitinated by Smad-specific E3 ubiquitin ligase SMURF1, leading to its degradation via proteasome Smad1 variants are associated with susceptibility to pulmonary hypertension that is characterized by plexiform lesions of proliferating endothelial cells in pulmonary arterioles. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies.

Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphoserine 463/465 from the C-terminal region of human Smad1.

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany