AbFlex^® NRAS antibody (rAb)

Application Notes

Applications Validated by Active Motif:
IP: 4 µg per IP
WB*: 0.5-2 µg/ml
DB:1µg/µl

For optimal results, we recommend the addition of 0.05% Tween 20 to all blocking solutions to reduce background. Individual optimization may be required.

Immunogen

This information is proprietary to Active Motif and/or its suppliers.

Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis Tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.

AbFlex® NRAS antibody was expressed as full-length IgG with rabbit immunoglobulin heavy and light chains in mammalian 293 cells. The antibody was directly labeled with biotin using the biotin ligase, BirA.

Oncogenic mutations in Ras genes have been found in about 30% of human cancers. This family of genes is involved in cell signaling pathways that control cell growth and death. The human Ras gene family consists of three identified members H-, K- and NRAS encoding proteins of 188-189 amino acids and 21,000 (p21) daltons. The NRAS oncogene encodes a membrane protein that shuttles between the Golgi apparatus and the plasma membrane. This protein is activated by a guanine nucleotide-exchange factor and inactivated by a GTPase activating protein. Human H- and K-Ras are the homologues of v-H- and v-K-ras, sequences. Under normal circumstances the Ras-GTPase activity gradually converts GTP to GDP, but in oncogenic mutations Ras locks the protein into the active GTP conformation. This leads to a constant Ras gene and uncontrollable cell growth. Due to these intracellular relationship Ras antibodies have emerged as a valuable tool for target validation studies for cancer.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

• ChIP = Chromatin Immunoprecipitation;
• ChIP = ChIP Sequencing;
• CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
• CUT&Tag = Cleavage Under Targets and Tagmentation;
• DB = Dot Blot;
• ELISA = Enzyme-linked Immunosorbent Assay;
• EMSA = Electrophoretic Mobility Shift Assay
• FC = Flow Cytometry;
• ICC = Immunocytochemistry;
• IF = Immunofluorescence;
• IHC = Immunohistochemistry;
• IP = Immunoprecipitation;
• MeDIP = Methyl-DNA Immunoprecipitation;
• NEU = Neutralizing;
• WB = Western Blotting