Anti-p-MST1/2 (Thr183/180) Antibody, clone 1C2 ZooMAb^® Rabbit Monoclonal

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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.

Clone 1C2 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects MST1/MST2 phosphorylated on Threonine 183 (in MST1) and Threonine 180 (in MST2).

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphotheonine 183 from human MST1/STK4.

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells treated with Staurosporine.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected phospho-MST1 (Thr183)/MST2 (Thr180) in lysate from overnight serum starved HeLa cells treated Staurosporine (0.5 M; 6 h), but not in lysate from untreated cells.

Tested Applications

Affinity Binding Assay: A representative lot of this antibody bound phosphorylated MST1/MST2 peptide ((Thr183/180) with a KD of 4.4 x 10-6 in an affinity binding assay.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected p-MST1/2 (Thr183/180) in HeLa cells treated with Staurosporine.

Peptide Inhibition Assay:: Target band detection in HeLa cell lysate was prevented by pre-blocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

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

Serine/threonine-protein kinase 4/3 (UniProt: Q13043/Q13188; also known as EC: 2.7.11.1, Mammalian STE20-like protein kinase 1/2, MST-1/2, STE20-like kinase MST1/2, Serine/threonine-protein kinase Krs-2/1, STK4/3) is encoded by the STK4/3 (also known as KRS2/1, MST1/2) gene (Gene ID: 6789/6788) in human. MST1/2 are homodimeric, stress-activated, pro-apoptotic kinases that enter the nucleus following its caspase cleavage and induces chromatin condensation followed by internucleosomal DNA fragmentation. They are  ubiquitously expressed with higher expression observed in cancer tissues and its levels increase with the progression to malignant state. Their protein kinase domain is localized in residues 30-281 and 27-278, for MST1 and MST2, respectively.  They have an N-terminal catalytic domain and C-terminal regulatory region. They are activated by caspase-cleavage between amino acids 326-327 and 349-350 for MST1 and residues 322-323 for MST2.  Their caspase-cleaved form cycles between the nucleus and cytoplasm. Their full activation requires homodimerization and autophosphorylation at threonine 183 and 180, respectively for MST1 and MST2. Their activity is inhibited by the C-terminal non-catalytic region. They are KEY^® components of the Hippo signaling pathway, which plays a pivotal role in organ size control and acts as a tumor suppressor by restricting cell proliferation and promoting apoptosis. During apoptosis, MST1 phosphorylates serine 14 of histone H2B and phosphorylates FOXO3 upon oxidative stress that results in its NuCLEAR^™ translocation and initiation of cell death. Its phosphorylation at threonine 387 by AKT1, LEADS^™ to inhibition of its kinase activity, NuCLEAR^™ translocation and autophosphorylation at threonine 183. It also reduces its cleavage by caspases and its ability to phosphorylate FOXO3. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: (Ref.: Guennoun, A., et al. (2021). J. Clin. Immunol. 41(8); 1839-1852; Praskova, M., et al. (2004). Biochem. J. 381(Pt2); 453-462; Lee, KK., et al. (2001). J. Biol Chem. 276(22); 19276-19285).

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 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.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

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.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Affinity is a trademark of Mine Safety Appliances Co.

KEY is a registered trademark of Hanna Instruments

LEADS is a trademark of Compugen

NuCLEAR is a trademark of Sigma-Aldrich Co. LLC

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

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.