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AB325444

Human TNF alpha ELISA Kit, Chemiluminescent

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Human TNF alpha ELISA Kit, Chemiluminescent is a 55 min single-wash SimpleStep Ignite™ ELISA for the rapid quantitation of Human TNF alpha with higher sensitivity and broader measuring range.

- Single 30 min incubation and single-wash for results in less than an hour
- Chemiluminescent detection allows a broad dynamic range: 2.1 - 4500pg/ml with a sensitivity of 0.66pg/ml
- Validated on a number of sample types including cerebrospinal fluid (CSF)
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

TNFA, TNFSF2, TNF, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a

5 Images
Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 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.

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)

Interpolated concentrations of native TNF alpha in unstimulated versus stimulated PBMC cell culture supernatant samples. PBMC cells were stimulated with 1.5% PHA-M or vehicle control in 10F RPMI and incubated for 46 hours. The concentrations of TNF alpha were measured in duplicate and interpolated from the TNF alpha standard curves. Undiluted samples are PHA-M stimulated PBMC cell culture supernatant 10% and unstimulated PBMC cell culture supernatant 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was back calculated determined to be 7,265.71 pg/mL in PHA-M stimulated PBMC cell culture supernatant and undetectable in the unstimulated PBMC control.

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)

Dynamic range comparison between the Chemiluminescent Human TNF alpha SimpleStep ELISA and colorimetric Human TNF alpha SimpleStep ELISA (ab181421).

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)

Standard curve comparison between the Chemiluminescent Human TNF alpha SimpleStep ELISA and colorimetric Human TNF alpha SimpleStep ELISA (ab181421) in Sample Diluent NS. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit, Chemiluminescent (AB325444)

Interpolated concentrations of native TNF alpha in human PBMC cell culture supernatant samples. PBMC cells were treated with 1.5% PHA-M for 46 hours in 10F RPMI. The concentrations of TNF alpha were measured in duplicate, interpolated from the TNF alpha standard curve and corrected for sample dilution. Undiluted samples are PHA-M treated PBMC cell culture supernatant 10%. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2).

Key facts

Detection method

Luminescent

Sample types

Serum, Citrate plasma, EDTA Plasma, Heparin Plasma, Cell culture supernatant, Cerebral Spinal Fluid

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 0.66 pg/mL

Range

2.1 - 4500 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human TNF alpha ELISA Kit, Chemiluminescent (ab325444) is a rapid single-wash sandwich 55-min ELISA to measure Human TNF alpha in Serum, Citrate Plasma, EDTA Plasma, Heparin Plasma, Cell Culture Supernatant, Cerebrospinal Fluid with a sensitivity of 0.66 pg/ml.

How the assay works

Human TNF alpha ELISA Kit, Chemiluminescent (ab325444) SimpleStep ELISA® employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colormetric ELISAs.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.

Please refer to our protocol booklet for more details.

Human TNF alpha (ab325444) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature

2. Wash

3. Add prepared ChemiHRP development solution - incubate for 2 min

4. Read the results on a plate reader capable of luminescence detection

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-TNF alpha antibody [EPR18565-29] BSA and Azide free (Capture) ab242476

- Anti-TNF alpha antibody [EPR18565-33] BSA and Azide free (Detector) ab242978

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Precision

[ { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 2.5" }, { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 3.1" } ]
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Recovery

[ { "sample": "Serum", "range": "83 - 93 %", "average": "= 88" }, { "sample": "Citrate plasma", "range": "86 - 97 %", "average": "= 91" }, { "sample": "EDTA Plasma", "range": "95 - 104 %", "average": "= 100" }, { "sample": "Heparin Plasma", "range": "89 - 110 %", "average": "= 98" }, { "sample": "Cell culture supernatant", "range": "105 - 113 %", "average": "= 110" }, { "sample": "Cerebral Spinal Fluid", "range": "80 - 88 %", "average": "= 83" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

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Target data

Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Up-regulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed : 23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed : 16829952, PubMed : 22517918, PubMed : 23396208). Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance (By similarity). Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6 (PubMed : 12794819). Promotes osteoclastogenesis and therefore mediates bone resorption (By similarity).. The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.
See full target information TNF
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websiteProtocolBooklet
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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