LIGHT/TNFSF14 was detected in immersion fixed paraffin-embedded sections of human spleen using Goat Anti-Human LIGHT/TNFSF14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF664) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to splenocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Cell Proliferation Induced by LIGHT/TNFSF14 and Neutral-ization by Human LIGHT/ TNFSF14 Antibody.

Recombinant Human LIGHT/ TNFSF14 (Catalog # 664-LI) induces proliferation in the HUVEC human umbilical vein endothelial cells in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human LIGHT/ TNFSF14 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human LIGHT/ TNFSF14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF664). The ND₅₀ is typically 5-20 ng/mL.

Human LIGHT/TNFSF14 ELISA Standard Curve.

Recombinant Human LIGHT/TNFSF14 protein was serially diluted 2-fold and captured by Mouse Anti-Human LIGHT/TNFSF14 Monoclonal Antibody (MAB6642) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human LIGHT/TNFSF14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF664) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

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Applications for Human LIGHT/TNFSF14 Antibody

Application

Recommended Usage

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human LIGHT/TNFSF14 Monoclonal Antibody (Catalog # MAB6642).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human LIGHT/TNFSF14 DuoSet ELISA Kit (Catalog # DY664) for convenient development of a sandwich ELISA or the Human LIGHT/TNFSF14 Quantikine ELISA Kit (Catalog # DLIT00) for a complete optimized ELISA.

Immunohistochemistry

3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human spleen

Western Blot

0.1 µg/mL
Sample: Recombinant Human LIGHT/TNFSF14 (Catalog # 664-LI)

Neutralization

Measured by its ability to neutralize LIGHT/TNFSF14-induced proliferation in the HUVEC human umbilical vein endothelial cells. Conn, G. et al. (1990) Proc. Natl. Acad. Sci USA 87:1323. The Neutralization Dose (ND₅₀) is typically 5-20 ng/mL in the presence of 3 ng/mL Recombinant Human LIGHT/TNFSF14.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: LIGHT/TNFSF14

Human LIGHT is a type II membrane protein that is a member of the TNF superfamily. LIGHT is an acronym which stands for "is homologous to lymphotoxins, exhibits inducible expression, and competes with HSV glycoprotein D for HVEM, a receptor expressed by T lymphocytes". LIGHT has also been called HVEM-L and LT-gamma. Under the new TNF nomenclature, it is called TNFSF14. LIGHT is a 240 amino acid (aa) protein that contains a 37 aa cytoplasmic domain, a 22 aa transmembrane region, and a 181 aa extracellular domain. Similar to other TNF ligand family members, LIGHT is predicted to assemble as a homotrimer. LIGHT is produced by activated T cells and was first identified by its ability to compete with HSV glycoprotein D for HVEM binding. LIGHT has also been shown to bind to the lymphotoxin beta receptor (LT betaR) and the decoy receptor (DcR3/TR6). LIGHT overexpression in tumor cells induces apoptosis, which can be enhanced by IFN-gamma. The full roles of LIGHT remain to be elucidated.

References

Mauri, D.N. et al. (1998) Immunity 8:21.
Zhai, Y. et al. (1998) J. Clin. Invest. 102:1142.
Harrop, J.A. et al. (1998) J. Biol. Chem. 273:27548.
Yu, K-Y. et al. (1999) J. Biol. Chem. 274:13733.

Long Name

TNF Ligand Superfamily Member 14

Alternate Names

CD258, TNFSF14

Entrez Gene IDs

8740 (Human); 50930 (Mouse)

Gene Symbol

TNFSF14

UniProt

O43557

Additional LIGHT/TNFSF14 Products

Product Documents for Human LIGHT/TNFSF14 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human LIGHT/TNFSF14 Antibody

For research use only

Citations
Reviews

Human LIGHT/TNFSF14 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF664

Key Product Details

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Endotoxin Level

Scientific Data Images for Human LIGHT/TNFSF14 Antibody

LIGHT/TNFSF14 in Human Spleen.