DESCRIPTION

Syntaxins are a family of transmembrane proteins that belong to the SNARE complex. In conjunction with other SNAREs and the cytoplasmic NSF and SNAP proteins, syntaxins mediate vesicle fusion in diverse vesicular transport processes along the exocytic and endocytic pathways. Syntaxin 1A (STX1A) and syntaxin 1B (STX1B), two closely related isoforms of syntaxin 1, are involved in synaptic vesicle docking and fusion during neurotransmitter release. STX1A and STX1B are expressed in neurons and are localized on the neuronal plasma membrane. Human STX1B shares 100% amino acid sequence identity with mouse and rat STX1B within the cytoplasmic domain.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the 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	1,717.2	37.7	2.2
2	8	452.5	10.3	2.3
3	8	242.5	9.2	3.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,765.3	77.4	4.4
2	16	450.6	13.1	2.9
3	16	239.6	7.5	3.1

RECOVERY

The recovery of human/mouse STX1B spiked to three different levels throughout the range of the assay in tissue lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Tissue lysate	1:6,400	105	99-109
	1:12,800	105	102-109

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 tissue lysate was initially diluted 1:800.)

		Tissue lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	125
	Range (%)	120-129
1:8	Average% of Expected	96
	Range (%)	94-97
1:16	Average% of Expected	116
	Range (%)	99-134

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human/Mouse STX1B One-Step ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

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