Anti-ICA 512/RESP18 Antibody, clone 1348-D29-1

Receptor-type tyrosine-protein phosphatase-like N (UniProt: P16849; also known as R-PTP-N, Islet cell antigen 512, ICA 512, Islet cell autoantigen 3, PTP IA-2) is encoded by the PTPRN (also known as ICA3, ICA512) gene (Gene ID: 5798) in human. ICA 512, a tyrosine phosphatase like intrinsic membrane protein, is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-34), which is subsequently cleaved off to produce the mature form that contains a lumenal domain (aa 35-575), a transmembrane domain (aa 576-600), and a cytoplasmic domain (aa 601-979). Its lumenal region contains the RESP18 homology domain that is localized in amino acids 35-131. The mature form can undergo cleavage to generate an N-terminal fragment (ICA 512-NTF), a transmembrane fragment (ICA 512-TMF), and a cleaved cytosolic fragment (ICA 512-CCF). ICA 512 expression is restricted to neuroendocrine cells and is detected in the pancreas, brain, and pituitary. It is involved in the biogenesis and turnover of insulin secretory granules (SGs) in pancreatic islet β-cells . It plays a role in insulin secretion in response to glucose. It is required for the accumulation of normal levels of insulin-containing vesicles and preventing their degradation. It plays a role in insulin secretion in response to glucose stimuli. It is also required for normal accumulation of secretory vesicles in hippocampus and pituitary. ICA 512-TMF is reported to regulate dynamics and exocytosis of insulin SGs. ICA 512-CCF is shown to translocate to the nucleus and promotes the expression of insulin and other granule-related genes. It enhances pancreatic b-cell proliferation by converging with signaling by STAT5B and STAT3. ICA 512-NTF containing the RESP18HD domain encodes for an intrinsically disordered region that acts as a condensing factor for the reversible aggregation of insulin and other β-cell proteins in a pH and zinc-regulated manner. (Ref.: Toledo, PL., et al. (2019). J. Biol. Chem. 294(21); 8564-8576; Mziaut, H., et al. (2008). Proc. Natl. Acad. Sci. USA. 105(2); 674-679; Trajkovski, M., et al. (2008). J. Biol. Chem. 283(48); 33719-33729).

Clone 1348-D29-1 is a mouse monoclonal antibody that detects ICA 512/RESP18. It targets an epitope within the RESP18 homology domain within the N-terminal fragment.

GST-tagged recombinant protein fragment corresponding to the mature N-terminal fragment (without the signal peptide) of human ICA 512.

Quality Control Testing

Evaluated by Western Blotting in lysate from EndoC βH1 cells.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected ICA 512/RESP18 in EndoC βH1 cells, but not in cells subjected to RNAi knockdown.

Tested Applications

Immunofluorescence Analysis: A representative lot detected ICA 512/RESP18 in Immunofluorescence applications (Toledo, P.L., et al. (2019). J Biol Chem. 294(21):8564-8576).

Western Blotting Analysis: A representative lot detected ICA 512/RESP18 in Western Blotting application (Toledo, P.L., et al. (2019). J Biol Chem. 294(21):8564-8576).

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

Purified mouse monoclonal antibody IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

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

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