Nori Canine IL-14A/TXLNA ELISA Kit Summary

Alternative names for IL-14A: Interleukin 14A, IL14A, Alpha-taxilin, TXLNA, TXLN

Alternative names for canine: Dog

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Interleukin-14 alpha (IL-14A), also known as alpha-taxilin (encoded by the TXLNA gene) [1], is a high-molecular-weight B-cell growth factor (HMW-BCGF) that promotes the proliferation of normal and cancerous B-cells [2]. It is involved in immune responses and is implicated in the pathogenesis of autoimmune diseases like SLE and Sjogren’s syndrome. IL-14A induces B-cell proliferation, inhibits antibody secretion by inhibiting differentiation into antibody-secreting plasma cells, and expands selected B-cell subgroups. IL-14A is involved in Ca2+-dependent exocytosis in neuroendocrine cells [3]. This interleukin is produced mainly by T cells and certain malignant B cells. IL-14A has significant clinical value as a predictive biomarker and a key mediator in autoimmune disease pathogenesis. Serum levels of IL-14α can serve as a biomarker to monitor responses to immunotherapy in patients with advanced solid cancers. IL-14α is considered a putative biomarker for the stratification and diagnosis of Sjögren’s disease (SjD) and related dry eye conditions. Patients with primary and secondary Sjögren’s syndrome express significantly higher levels of IL-14α in their peripheral blood compared to healthy individuals. Elevated levels are found in aggressive B-cell non-Hodgkin’s lymphoma, Sjögren’s syndrome, and Systemic Lupus Erythematosus (SLE). Early changes in serum IL-14α levels (measured after two cycles of anti-PD-1 therapy like pembrolizumab) can predict patient survival and treatment response. While the precise nature of interleukin-14 remains under study, the alpha subunit has been identified as a critical factor in the development of B-cell-related disorders.

References

1. Nogami S, et al. (2004). Biochem. Biophys. Res. Commun. 319 (3): 936–43.
2. Ambrus JL, et al.(1993). Proc. Natl. Acad. Sci. U.S.A. 90 (13): 6330–4.
3. Nogami S, et al. (2003). Genes Cells. 8 (1): 17–28.