Application

Research CategoryNeuroscience

Anti-BRAP, clone 3C18 Antibody, Cat. No. MABN801, is a highly specific mouse monoclonal antibody that targets BRAP and has been tested in Immunocytochemistry and Western Blotting.

Immunocytochemistry Analysis: 5 µg/mL from a representative lot detected EGFP-tagged mouse BRAP in transfected COS-7 cells (Courtesy of Yuanyi Feng, Ph.D., Northwestern University, USA).

Western Blotting Analysis: 0.5 µg/mL from a representative lot detected exogenously overexpressed mouse BRAP in 25-50 µg of lysate from transfected HEK293T cells (Courtesy of Yuanyi Feng, Ph.D., Northwestern University, USA).

Immunocytochemistry Analysis: A representative lot detected BRAP immunoreactivity co-localized with that of LIS1 beneath the plasma membrane by fluorescent immunocytochemistry staining of mouse embryonic fibroblasts (MEFs), 293FT and COS1 cells (Lanctot, A.A., et al. (2013). Dev. Cell. 25(3):241-255).

Western Blotting Analysis: A representative lot detected both the endogenous human BRAP in untransfected HEK293T as well as the exogenously expressed myc-tagged murine BRAP in transfected HEK293T cells (Lanctot, A.A., et al. (2013). Dev. Cell. 25(3):241-255).

Note: DO NOT HEAT SAMPLES PRIOR TO GEL LOADING. BRAP protein tends to form insoluble aggregates upon heating.

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.

General description

BRCA1-associated protein (UniProt Q99MP8; also known as BRAP2, IMP, Impedes mitogenic signal propagation) is encoded by the Brap gene (Gene ID 72399) in murine species. BRAP is a Ras effector that controls the sensitivity of MAPK signaling by negatively regulating the MAPK scaffold protein KSR1 (kinase suppressor of Ras 1), allowing adaptations and homeostasis in a complex tissue environment. In the developing CNS, mitogenic/morphogenic molecules from location-specific signaling centers form counteracting gradients and simultaneously guide orchestrated cell proliferation, neuronal differentiation, and tissue morphogenesis. Consistent with its role as a MAPK signaling threshold modulator, BRAP2 is shown to interact with both Nde1 and Lis1 to form a complex that provides proper buffer strength to set up the signaling threshold according to the local concentration of mitogens and morphogens. The Nde1-Lis1-BRAP complex helps prevent excessive activation of MAPK under high local mitogenic activity in the midline and promote the self-renewal of progenitors that are programmed to generate a large cohort of neurons, but are localized distant from the midline.

Immunogen

Recombinant murine BRAP.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Protein G purified.

Format: Purified

Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide

Quality

Evalulated by Western Blotting in mouse BRAP-expressing HEK293T cell lysate.

Western Blotting Analysis: 1 µg/mL of this antibody detected exogenously expressed mouse BRAP in 20 µg of lysate from transfected HEK293T cells.

Specificity

Clone 3C18 specifically detected the target band in wild-type, but not Brap-knockout MEF lysate. Clone 3C18 epitope region is absent in human spliced isoform 2 and mouse spliced isoform 3 (UniProt Q7Z569-2 and Q99MP8-3).

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~67 kDa observed. 67.31 kDa (human isoform 1), 66.99/63.95 kDa (mouse isoform 1/2; isoform alpha/beta), 65.58 kDa (rat) calculated. Uncharacterized bands may be observed in some lysate(s).