ANTI-RAGA/B CLONE 1H7.1

General description

Ras-related GTP-binding protein A (UniProt Q7L523; also known as Adenovirus E3 14.7 kDa-interacting protein 1, FIP-1. Rag A, RagA) is encoded by the RRAGA gene (Gene ID 10670) in human. Mammalian cells express four Rag GTPases that mediate amino acid sufficiency-dependent mTORC1 activation to support cell growth and proliferation. Rag GTPases assemble into heterodimers containing GTP-loaded RagA or RagB partnered with GDP-loaded RagC or RagD. Promoted by signal from an upstream amino acid-responsive supercomplex, the active Rag heterodimer accumulates on the cytoplasmic surface of the lysosome, where it recruits mTORC1 and initiates its activation. The pentameric Ragulator complex tethers Rag heterodimers to the lysosomal membrane and acts as RagA/B guanine nucleotide exchange factor (GEF), while the heterotrimeric GATOR1 complex and the heterodimeric FNIP-Folliculin complex function as RagA/B and RagC/D GTPase activating protein (GAP), respectively. Both lysosomal amino acid sensors, such as the multisubunit vacuolar ATPase (v-ATPase) and lysosomal amino acid transporter(s), and cytoplasmic amino acid sensors, such as the leucyl-tRNA synthetase (LeuRS), are part of the upstream amino acid-responsive supercomplex.

Specificity

Clone 1H7.1 targets an epitope within the C-terminal end sequence highly conserved between RagA and RagB.

Immunogen

KLH-conjugated linear peptide corresponding to the C-terminal end sequence of human/mouse/rat RagA.

Application

Detect RagA and RagB using this mouse monoclonal Anti-RagA/B, clone 1H7.1, Cat. No. MABS1182, validated for use in ELISA, Immunocytochemistry, and Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected exogenously overexpressed human RagA in 1 µg of lysate from transfected HEK293T cells.

Immunocytochemistry Analysis: A 1:50 dilution from a representative lot detected RagA/B immunoreactivity in HUVECs.

ELISA Analysis: Clone 1H7.1 hybridoma culture supernatant detected exogenously overexpressed human RagA and RagB in lysates (100 ng/well) from transfected HEK293T cells.

Quality

Evaluated by Western Blotting in HEK293.

Western Blotting Analysis: A 1:5,000 dilution of this antibody detected RagA in 10 µg of HEK293 cell lysate.

Target description

~36 kDa observed. 36.57 kDa (human/mouse/rat/bovine RagA), 43.25/40.17 kDa (human RagB Long/Short isoform), 43.19 kDa (mouse RagB), 43.19/40.11 (rat RagB Long/Short isoform) calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

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

Other Notes

Concentration: Please refer to lot specific datasheet.

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.