LTA ELISA Kit

Catalog No. ABIN1112685

Product Details LTA ELISA Kit

Target: LTA (Lymphotoxin-alpha (LTA))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Minimum Detection Limit: 62.5 pg/mL

Application: ELISA

Purpose: For quantitative detection of TNF in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Sample Type: Cell Culture Supernatant, Serum, Tissue Lysate

Analytical Method: Quantitative

Sensitivity: < 5 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human TNFbeta antibody 2. Lyophilized human TNFbeta standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-human TNF beta antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-P- Cadherin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-TNF polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the TNF amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of TNF can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate or body fluids, cell culture supernate: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin or EDTA as the anticoagulant. Centrifuge for 15 min at 1000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for LTA

Target: LTA (Lymphotoxin-alpha (LTA))

Alternative Name: TNFbeta (LTA Products)

Synonyms: LT ELISA Kit, TNFB ELISA Kit, TNFSF1 ELISA Kit, LT-[a] ELISA Kit, LT-alpha ELISA Kit, LT[a] ELISA Kit, LTalpha ELISA Kit, Ltx ELISA Kit, TNF-beta ELISA Kit, Tnfb ELISA Kit, Tnfsf1b ELISA Kit, hlb382 ELISA Kit, lymphotoxin alpha ELISA Kit, lymphotoxin A ELISA Kit, LTA ELISA Kit, Lta ELISA Kit

Target Type: Chemical

Background: Tumor necrosis factor-beta (TNF-beta), previously called lymphotoxin-alpha(LTA), is a member of the tumor necrosis factor family , is a cytokine produced by lymphocytes. It is a glycoprotein with a relative molecular mass (Mr) of 60,000-70,000. LTA is highly inducible, secreted, and exists as homotrimeric molecule. It forms heterotrimers with lymphotoxin-beta which anchors lymphotoxin-alpha to the cell surface. LTA mediates a large variety of inflammatory, immunostimulatory, and antiviral responses. It is also involved in the formation of secondary lymphoid organs during development and plays a role in apoptosis.

Pathways: Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process