Anti-p-ATF-2-Thr69/71 Antibody, clone 1H23 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 1H23 is a ZooMAb^® Rabbit recombinant monoclonal antibody that specifically detects ATF-2 phosphorylated on threonine 69 and 71.

KLH-conjugated linear peptide corresponding to 14 amino acids surrounding phosphothreonine 69/71 from the N-terminal region of human ATF-2.

Quality Control Testing

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

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected ATF2 phosphorylated at Threonine 69 and 71 in lysate from HeLa cells, overnight serum starved and treated with Anisomycin (25 g/mL; 30 min), but not in lysate from untreated cells.

Tested applications

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected a construct containing MBP-tagged recombinant fragment of ATF-2 phosphorylated on threonine 69/71, but did not detect in non-phosphorylated construct. (Phospho- ATF-2 construct: Courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine).

Peptide Inhibition Assay Analysis: Target band detection in lysate from HeLa cells treated with Anisomycin 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-ATF-2-Thr69/71 peptide with a KD of 2.1 x 10-8 in an affinity binding assay.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Phospho-ATF-2-Thr69/71 in human lung cancer tissue sections.

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

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

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected ATF2 phosphorylated at Threonine 69 and 71 in lysate from HeLa cells, overnight serum starved and treated with Anisomycin (25 g/mL; 30 min), but not in lysate from untreated cells.

Cyclic AMP-dependent transcription factor ATF-2 (UniProt: P15336; also known as cAMP-dependent transcription factor ATF-2, Activating transcription factor 2, Cyclic AMP-responsive element-binding protein 2, CREB-2, cAMP-responsive element-binding protein 2, HB16, cAMP response element-binding protein CRE-BP1) is encoded by the ATF2 (also known as CREB2, CREBP1) gene (Gene ID: 1386) in human. ATF2 is a member of the activating protein-1 (AP1) transcription factor family that regulates the transcription of various genes, including those involved in anti-apoptosis, cell growth, and DNA damage response. Dependent on its binding partner, it binds to cAMP response element (CRE) consensus sequences (5′-TGACGTCA-3′) or to AP-1 consensus sequences (5′-TGACTCA-3′). ATF2 is reported to shuttle between the cytoplasm and the nucleus and its heterodimerization with Jun is essential for its nuclear localization. It is localized in the cytoplasm under conditions of cellular stress and in disease states. It can also localize at the mitochondrial outer membrane in response to genotoxic stress. In the cytoplasm, it is shown to interact with and perturbs hexokinase 1 and VDAC1-containing complexes at the mitochondrial outer membrane and impair mitochondrial membrane potential that leads to mitochondrial leakage and promoting cell death. Under basal conditions, ATF2 is auto-inhibited through intramolecular contact between the zinc finger (aa 25-49) and transactivation domain (TAD)-containing N-terminal domain and C-terminal basic leucine zipper (bZIP; aa 352-415) domain. Following activation by kinases that phosphorylate the N-terminal region, it is activated and translocates to the nucleus to modulate gene expression. Its phosphorylation at threonine 69 and 71 by MAP kinases, in response to external stimuli, increases its transcriptional activity. This ZooMAbZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Bhoumik, A., et al. (2005). Mol. Cell. 18(5); 577-587; Ouwens, DM., et al. (2002). EMBO J. 21(14); 3782-3793).

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.

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.

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.