Analysis Note

ControlIncluded Positive Antigen Control: Catalog # 12-301, non-stimulated A431 lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Research Sub CategoryMAP Kinases

Detect Ha-Ras using this Anti-Ha-Ras Antibody, clone MC57 validated for use in WB.

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

Ras proteins are small GTP-binding proteins which, unlike the heterotrimeric G-proteins, contain all GTPase and effector functions within a single polypeptide. At least three isoforms of Ras exist, Ki-Ras, Ha-Ras and N-Ras, with distinct expression patterns but similar signaling activity. Ras is palmitoylated and farnesylated at the carboxy terminus, anchoring it in the membrane. In resting cells, Ras is loaded with GDP, and is activated subsequent to growth factor stimulation of receptors, which recruit Ras Guanine nucleotide Exchange Factors to the plane of the membrane. Proximity of exchange factors to the Ras proteins causes release of GDP, and its replacement by GTP. In its GTP-bound form, Ras binds several proteins, including Raf, RalGDS and PI3 Kinase. Inactivation of Ras occurs by GTP hydrolysis, which is greatly accelerated by RasGAP or NF-1, two known Ras GTPase Activating Proteins. It is possible to assay for Ras activation by incubation of lysates with the Ras-binding domain of Raf-1, which selectively binds to Ras:GTP.

Immunogen

GST fusion protein corresponding to full length human Ha-Ras. Clone MC57.

Linkage

Replaces: 05-775

Other Notes

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

Physical form

Cultured supernantant containing 0.05% sodium azide.

Quality

Western Blot Analysis: A 1:1000 dilution of this antibody detected Ha-Ras in RIPA lysates from A431 cells.

Specificity

Predicted to cross-react with mouse and rat based on sequence homology.

Recognizes Ha-Ras, Mr ~21 kDa. Does not cross react with Cdc42, Rho or Rac1.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~21 kDa