DESCRIPTION

RAF1, also known as Raf-1 proto-oncogene serine/threonine kinase, plays pivotal roles in the MAPK/ERK signaling pathway. It is activated by growth factor receptors and other extracellular stimuli, promoting cell proliferation, differentiation, and survival. Upon activation, RAF1 phosphorylates and activates MEK, which in turn activates ERK. Abnormal activation of RAF1 is implicated in various cancers. Mutations or overexpression of RAF1 can lead to constitutive activation of the MAPK pathway, driving cell proliferation and tumorigenesis. For example, RAF1 is involved in the development of multiple tumors, such as non-small cell lung cancer.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns solution yellow. The color intensity is proportional to the quantity of bound protein which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	604.1	29.1	4.8
2	8	177.4	1.7	1.0
3	8	56.9	1.4	2.5

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	609.7	29.5	4.8
2	16	169.8	8.6	5.1
3	16	53.8	3.9	7.3

RECOVERY

The recovery of human NCX1 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:32	97	90-100
	1:64	100	96-103

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The cell lysate was initially diluted 1:4.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	96
	Range (%)	78-113
1:8	Average% of Expected	89
	Range (%)	79-98
1:16	Average% of Expected	90
	Range (%)	74-105

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human RAF1 Sandwich ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
• Payment: T/T 100% in advance; PayPal; Alibaba Trade Assurance
• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)