Anti-LIGHT/Tnfsf14 Antibody Picoband™

Product Name

Anti-LIGHT/Tnfsf14 Antibody Picoband™

View all LIGHT/TNFSF14 Antibodies

SKU/Catalog Number

A03516-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-LIGHT/Tnfsf14 Antibody Picoband™ catalog # A03516-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-LIGHT/Tnfsf14 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03516-2)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse LIGHT/Tnfsf14 recombinant protein (Position: M1-V239).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A03516-2 is reactive to TNFSF14 in Mouse, Rat

Applications

A03516-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

30 kDa

Calculated molecular weight

26.35kDa

Background of LIGHT/TNFSF14

Tumor necrosis factor ligand superfamily member 14 is a protein that in humans is encoded by the TNFSF14 gene. TNFSF14 has also been designated as CD258, as well as LIGHT. It was mapped on chromosome 19p13.3. The protein encoded by this gene is a member of the tumor necrosis factor(TNF) ligand family. This protein may function as a costimulatory factor for the activation of lymphoid cells and as a deterrent to infection by herpesvirus. It has been shown to stimulate the proliferation of T cells, and trigger apoptosis of various tumor cells. This protein is also reported to prevent tumor necrosis factor alpha mediated apoptosis in primary hepatocyte. Two alternatively spliced transcript variant encoding distinct isoforms have been reported.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Mouse
Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Mouse, Rat
Flow Cytometry, 1-3 µg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5 µg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of LIGHT/Tnfsf14 using anti-LIGHT/Tnfsf14 antibody (A03516-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: mouse thymus tissue lysates,
Lane 2: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-LIGHT/Tnfsf14 antigen affinity purified polyclonal antibody (Catalog # A03516-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for LIGHT/Tnfsf14 at approximately 30 kDa. The expected band size for LIGHT/Tnfsf14 is at 26 kDa.

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Figure 2. IHC analysis of LIGHT/Tnfsf14 using anti-LIGHT/Tnfsf14 antibody (A03516-2).
LIGHT/Tnfsf14 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LIGHT/Tnfsf14 Antibody (A03516-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of LIGHT/Tnfsf14 using anti-LIGHT/Tnfsf14 antibody (A03516-2).
LIGHT/Tnfsf14 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LIGHT/Tnfsf14 Antibody (A03516-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of ANA-1 cells using anti-LIGHT/Tnfsf14 antibody (A03516-2).
Overlay histogram showing ANA-1 cells stained with A03516-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-LIGHT/Tnfsf14 Antibody (A03516-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For TNFSF14 (Source: Uniprot.org, NCBI)

Alternative Names

CD258 antigen; CD258; delta transmembrane LIGHT; Herpes virus entry mediator ligand; Herpesvirus entry mediator ligand; HVEM-L; HVEMLherpesvirus entry mediator-ligand; ligand for herpesvirus entry mediator; LIGHT; LIGHTherpesvirus entry mediator A; LTg; TNFSF14; TR2; tumor necrosis factor (ligand) superfamily, member 14; tumor necrosis factor ligand superfamily member 14; tumor necrosis factor receptor-like 2; tumor necrosis factor superfamily member LIGHT TNFSF14 CD258, HVEML, LIGHT, LTg TNF superfamily member 14 tumor necrosis factor ligand superfamily member 14|herpesvirus entry mediator ligand|tumor necrosis factor (ligand) superfamily, member 14|tumor necrosis factor ligand 1D|tumor necrosis factor superfamily member 14

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In this infographic, you will see the following information for TNFSF14: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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