Human TNF beta ELISA Kit (ab229202)
- Datasheet
- References
- Protocols
Overview
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Product name
Human TNF beta ELISA Kit
See all LTA kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% PBMC s/n 9 2.5% Inter-assay Sample n Mean SD CV% PBMC s/n 3 3.3% -
Sample type
Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
1.84 pg/ml -
Range
16 pg/ml - 1000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 101 98% - 103% Serum 82 82% - 83% Hep Plasma 87 87% - 89% EDTA Plasma 81 80% - 82% Cit plasma 80 79% - 81% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human, Rhesus monkey -
Product overview
Human TNF beta in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF beta protein in human serum, plasma and cell culture supernatants.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Tumor necrosis factor-beta, or Lymphotoxin-alpha and tumor necrosis factor ligand superfamily member 1, is a cytokine belonging to the tumor necrosis factor family and is encoded by the LTA gene. TNF-beta is a cytotoxic protein that is inducibly expressed from lymphocytes and has anti-proliferative activity, including initiating the cellular destruction of tumor cell lines. After secretion, TNF-beta forms a soluble homo-trimer or a membrane associated hetero-trimer with lymphotoxin-beta, which then anchors TNF-beta to the cellular surface. The heterotrimeric form binds to various receptors to activate the NF-kB pathway, which promotes immune regulation
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human TNF beta Capture Antibody 1 x 600µl 10X Human TNF beta Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent CPR 1 x 6ml Human TNF beta Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Cytokine that in its homotrimeric form binds to TNFRSF1A/TNFR1, TNFRSF1B/TNFBR and TNFRSF14/HVEM. In its heterotrimeric form with LTB binds to TNFRSF3/LTBR. Lymphotoxin is produced by lymphocytes and cytotoxic for a wide range of tumor cells in vitro and in vivo. -
Involvement in disease
Genetic variations in LTA are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). -
Sequence similarities
Belongs to the tumor necrosis factor family. -
Cellular localization
Secreted. Membrane. The homotrimer is secreted. The heterotrimer is membrane-associated. - Information by UniProt
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Alternative names
- LTalpha
- DAMA-25N12.13-004
- DIF
see all -
Database links
- Entrez Gene: 4049 Human
- Entrez Gene: 715425 Rhesus monkey
- Omim: 153440 Human
- SwissProt: P01374 Human
- SwissProt: Q5TM20 Rhesus monkey
- Unigene: 36 Human
Associated products
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Alternative Versions
Applications
Our Abpromise guarantee covers the use of ab229202 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Other - Human TNF beta ELISA Kit (ab229202)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human TNF beta standard curve in Sample Diluent NS.The TNF beta standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of spiked TNF beta in human serum and plasma samples.The concentrations of TNF beta were measured in duplicates, interpolated from the TNF beta standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 25%, plasma (heparin) 25% and plasma (EDTA) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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Interpolated concentrations of native TNF beta in human and Rhesus Macaque PBMC cell culture supernatants.PBMCs were either stimulated with 1.5% PHA or unstimulated for 48 hours. The concentrations of TNF beta were measured in duplicate and interpolated from the TNF beta standard curve and corrected for sample dilution. Undiluted samples are as follows: Stimulated human PBMC supernatant 25% and stimulated Rhesus Macaque PBMC supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF beta concentration was determined to be 1,660 pg/mL in the stimulated human PBMC supernatant, 1,940 pg/mL in the Rhesus Macaque stimulated PBMC supernatant, and undetectable in the unstimulated human PBMC supernatant (not shown).
Protocols
References
ab229202 has not yet been referenced specifically in any publications.
