Application

Western Blotting Analysis: A representative lot detected phospho RasGRP2 (pSer554) in a kinase assay with GST-Rasgrp2 (amino acids 1-374) and PKA, Forskolin and SKF81279 stimulated mouse striatal muscle slices, COS-7 cells transfected with EGFP-Rasgrp2 and Myc-PKA, nucleus accumbens (NAc) from mice treated with cocaine, Nucleus accu (Nagai, T., et. al. (2016). Neuron. 89(3):550-65).

Western Blotting Analysis: A representative lot detected RAS guanyl-releasing protein 2 purified from GST-Rasgrp2 transfected COS-7 cells and phosphorylated with GST-PKA catalytic subunit alpha, Forskolin stimulated mouse striatal muscle slices (Courtesy of Dr. Kozo Kaibuchi at Nagoya University, Japan).

Anti-phospho RasGRP2 (Ser554), Cat. No. ABS2095, is a rabbit polyclonal antibody that detects RAS guanyl-releasing protein 2 phosphorylated on Ser554 and has been tested for use in Western Blotting.

General description

RAS guanyl-releasing protein 2 (UniProt: Q9QUG9; also known as Calcium and DAG-regulated guanine nucleotide exchange factor I, RasGRP2, CalDAG-GEFI, F25B3.3 kinase-like protein) is encoded by the Rasgrp2 gene (Gene ID: 19395) in murine species. RasGRP proteins are considered as activators of Ras and other related small GTPases by virtue of functioning as guanine nucleotide exchange factors (GEFs). RasGRP-1 through -4 share many structural domains, however, they display subtle differences between each of the different family members. They are expressed in distinct patterns, such as in different cells of the hematopoietic system and in the brain. RasGRP2′s expression is restricted to platelets and their precursors, megakaryocytes, and neutrophils. RasGRP2 functions as a calcium- and DAG-regulated nucleotide exchange factor specifically activating Rap through the exchange of bound GDP for GTP. It may also activate other GTPases such as RRAS, RRAS2, NRAS, KRAS, but not HRAS. It is also reported to function in aggregation of platelets and adhesion of T-lymphocytes and neutrophils, probably through inside-out integrin activation. RaGRP2 is found both in the cytosol and associated with membranes. It is shown to be enriched at juxtamembrane areas and membrane ruffles. Three isoforms of RasGRP2 have been reported that are generated by alternative splicing. It contains two EF-hand domains, one N-terminal Ras-GEF domain, one phorbol-ester/DAG-type zinc finger domain, and one Ras-GEF domain. RasGRP2 deficient animals exhibit bleeding problems due to defects in platelets aggregation and degranulation. (Ref.: Olga Ksionda, O et al. (2013). Front Biol (Beijing). 8(5): 508 532).

Immunogen

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding Ser554 from the C-termnal region of murine RAS guanyl-releasing protein 2.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Format: Purified

Quality

Evaluated by Western Blotting with RAS guanyl-releasing protein 2 purified from GST-RasGRP2 transfected COS-7 cells and phosphorylated on Ser554 by GST-PKA calalytic subunit alpha (PKAc alpha).

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected phospho RasGRP2 (Ser554) purified from GST-RasGRP2 transfected COS-7 cells and phosphorylated on Ser554 by GST-PKA catalytic subunit alpha (PKAc alpha).

Specificity

This rabbit polyclonal antibody detects RAS guanyl-releasing protein 2 phosphorylated on Ser554 in various mammalian species. It targets an epitope within 11 amino acids surrounding Ser554 from the C-terminal region.

Target description

~90 kDa observed; 69.45 kda calculated. Uncharacterized bands may be observed in some lysate(s).