ANTI-GSDME/DFN5 CLONE 1H7 ZOOMAB« RABBIT MONOCLONAL (C005B-260751)

General description

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.

Specificity

Clone 1H7 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Gasdermin-E (GSDME). It targets an epitope within 20 amino acids from the N-terminal region.

Immunogen

KLH-conjugated linear peptide corresponding to 20 amino acids from the N-terminal region of human Gasdermin E (GSDME).

Application

Quality Control Testing

Evaluated by Western Blotting in A549 cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected GSDME/DFN5 in A549 cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected GSDME/DFN5 in SH-SY5Y cell lysate.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected GSDME/DFN5 in Human small intestine tissue sections.

Flow Cytometry Analysis: 1 µg from a representative lot detected GSDME/DFN5 in one million A549 cells.

Western Blotting Analysis: 0.1 µg/mL from a representative lot detected GSDME/DFN5 in lysates from HeLa cells, HEK293 cells, HEK293 cells transfected with DFNA5, and HeLa treated with TNF and cyclohexamide. (Data courtesy of Dr Kohsuke Tsuchiya from Kanazawa University, Japan).

Affinity Binding Assay: A representative lot of this antibody bound GSDME/DFN5 peptide with a KD of 5.3 x 10-8 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.

Target description

Gasdermin-E (UniProt: O60443; also known as Inversely correlated with estrogen receptor expression 1, ICERE-1, Non-syndromic hearing impairment protein 5, GSDME) is encoded by the GSDME (also known as DFNA5, ICERE1) gene (Gene ID: 1687) in human. Gasdermin-E (GSDME) is a precursor of a pore-forming protein that converts non-inflammatory apoptosis to pyroptosis. The full-length protein is inactive and upon cleavage by caspase 3 or granzyme B at aspartic acid 270, the N-terminal moiety is released (aa 1-270) that binds to membrane, homooligomerizes, and forms pores of 10 to 15 nm inner diameter, triggering pyroptosis. The N-terminal GSDME is a multi-pass membrane protein whose interaction with the C-terminal domain is important for autoinhibition in the absence of an activation signal.  GSDME expression is observed in cochlea and placenta. Lower levels of expression are reported in heart, brain, lung, liver, kidney, pancreas, and skeletal muscle. GSDME is also known to mediate secondary necrosis downstream of the mitochondrial apoptotic pathway and caspase-3 activation as well as in response to viral agents. It can suppress tumors by mediating granzyme-mediated pyroptosis in target cells of natural killer (NK) cells. It may also play a role in the p53/TP53-regulated cellular response to DNA damage. Three isoforms of GSDME have been described that are produced by alternative splicing. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: Zhang, Z., et al. (2020). Nature. 579(7799); 415-420; Wang, Y., et al. (2017). Nature. 547(7661); 99-103; Masuda, Y., et al. (2006). J. Hum. Genet. 51(8); 652-664).

Physical form

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.

Reconstitution

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.

Storage and Stability

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.

Other Notes

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

Legal Information

Affinity is a trademark of Mine Safety Appliances Co.

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

Disclaimer

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.