Abcam, ab134640, Ras GTPase ELISA Kit (Chemiluminescent)

Size: 1 x 96Tests
Ras GTPase ELISA Kit (Chemiluminescent) is a sandwich ELISA designed to quantify Ras GTPase (Chemiluminescent) with a sensitivity of 3000 ng/well. - Colorimetric sandwich ELISA - 450 nm readout - works on any plate reader - Wide dynamic range - quantifies 3000 - 25000 ng/well
Key facts
Detection method:Luminescent,
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Mouse, Human,
Assay type:Sandwich (quantitative),
Sensitivity:> 3000 ng/well,
Range:3000 - 25000 ng/well,
Assay time:4h 30m,
Assay Platform:Microplate

Product details:
Ras GTPase ELISA Kit (Chemiluminescent) ab134640 is a sandwich ELISA to measure Ras GTPase (Chemiluminescent) in cell culture extracts, tissue extracts with a sensitivity of 3000 ng/well.
How the assay works:
Ras GTPase ELISA Kits contain a Raf-RBD protein fused to GST that will be coated onto the provided 96-well (12 x 8 well plate), glutathione-coated plate. The activated Ras contained in cellular extract specifically binds to Raf-RBD, while inactive Ras does not bind. Bound Ras is detected by incubating with a primary antibody that detects H-Ras in mouse and H- & K-Ras in human samples. Addition of a secondary antibody conjugated to horseradish peroxidase (HRP) and developing solution provides a sensitive chemiluminescent readout that is easily quantified by luminescence. The 96-well plate with individual plate of 12 strips is suitable for manual use or high-throughput screening applications.
Assay Specificity
Ras GTPase ELISA Kit specifically detects activated H- and K-Ras in Human and H-Ras in rodent samples.
Ras GTPase ELISA Kit (Chemiluminescent) ab134640 protocol summary
1. Wash each well. Add sample to appropriate wells. Activated Ras binds Raf-RBD, inactive Ras does not. Incubate at room temperature.
2. Wash each well.
3. Add primary detection antibody. Incubate at room temperature.
4. Wash each well.
5. Add HRP-conjugated secondary antibody, which binds the primary antibody. Incubate at room temperature.
6. Wash each well. Add Chemiluminescent reagent.
7. Read using a luminometer.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Properties and Storage Information:
Shipped at conditions-MULTI, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The Ras protein also known by its alternate names such as KRAS NRAS and HRAS functions as a small GTPase which is a type of enzyme that binds to guanosine triphosphate (GTP) and can switch to an inactive form when it hydrolyzes GTP to GDP. Ras proteins are encoded by genes located in different chromosomes and typically have a molecular weight of about 21 kDa. These proteins are expressed in various tissues throughout the body where they play key roles in cellular signaling.
Biological function summary
Ras proteins are pivotal in regulating cell proliferation differentiation and survival. They are often part of larger protein complexes that facilitate signal transduction across cell membranes. These proteins function as binary molecular switches toggling between active (GTP-bound) and inactive (GDP-bound) states. Mutations in Ras proteins such as NRAS Q61R can lead the protein to assume permanently active conformations disrupting normal cellular signaling processes and contributing to oncogenesis.
Pathways
Ras proteins play significant roles in the MAPK/ERK and PI3K/AKT signaling pathways. Their activation leads to a cascade of phosphorylation events that ultimately regulate gene expression. Within these pathways Ras proteins interact with various molecules including RAF kinases and the PI3K protein further illustrating their complex roles in signal propagation. These pathways control many cellular processes including growth differentiation and survival illustrating how Ras proteins integrate multiple signals to modulate cellular outcomes.
Ras proteins are frequently implicated in various cancers notably pancreatic and colorectal cancers. Mutated forms of Ras such as KRAS G12V drive oncogenesis by promoting uncontrolled cell growth and division. In cancer pathology Ras often interacts with tumor suppressor proteins influencing the disease progression. Targeting Ras-mediated signaling pathways using approaches like pan-Ras inhibitors or chemiluminescence ELISAs represents an ongoing area of therapeutic research aimed at combating Ras-driven malignancies.