Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

TNF-alpha ELISA kit for Human

Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine™ (96 Tests). Quantitate Human TNF in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 1pg/ml. Cited in 10 publication(s).

Product Info Summary

SKU: EK0525
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

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Product Name

Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

View all TNF-alpha ELISA kits

SKU/Catalog Number

EK0525

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine™ (96 Tests). Quantitate Human TNF in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 1pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0525)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: V77-L233

Sensitivity

<1 pg/ml

Assay Range

15.6 pg/ml - 1,000 pg/ml

Cross Reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK0525 is reactive to TNF in Human samples

Validated Sample Types

cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

Application Guarantee

EK0525 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of TNF-alpha

Tumor necrosis factor-alpha (TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor (TNF) and Lymphotoxin (TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6.

Kit Components

Catalog Number Description Quantity
EK0525-CAP Anti-Human TNF Pre-coated 96-well strip microplate 1
EK0525-ST Human TNF Standard 2 vials, 10 ng/tube
EK0525-DA Human TNF Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015.631.262.51252505001000
O.D.0.0910.1580.1970.3200.5480.8971.5762.047

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect TNF-alpha, Dilution ratio of 1:1, concentration in serum is around 100pg/ml.

Some articles we found to cite concentrations of TNF-alpha in samples: 24812899 (Pubmed IDs).

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)16822011475186
Standard deviation0.765.4912.660.755.5511.9
CV (%)4.8%6.7%7.4%5.4%7.4%6.4%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of TNF-alpha in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 116151516150.53.3%
Sample 282917589846.297.4%
Sample 32012042152192097.463.5%
*number of samples for each test n=16.

Gene/Protein Information For TNF (Source: Uniprot.Org, NCBI)

Uniprot ID

P01375

Gene ID

7124

Gene Name

TNF

Full Name

Tumor necrosis factor

Weight

25.644kDa

Superfamily

Tumor necrosis factor family

Alternative Names

APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha TNF DIF-alpha, TNFA, TNFSF2, TNLG1F, TNF tumor necrosis factor tumor necrosis factor|APC1 protein|TNF, macrophage-derived|TNF, monocyte-derived|TNF-a|tumor necrosis factor ligand 1F|tumor necrosis factor ligand superfamily member 2|tumor necrosis factor-alpha

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on TNF, check out the TNF Infographic

TNF infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for TNF: database IDs, super-family, 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 us [email protected]

EK0525 has been cited in 10 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Inflammatory factor in donor liver and its effect on recipient myocardial injury after liver transplantation.

Inflammatory response of macrophages cultured with Helicobacter pylori strains was regulated by miR-155

Serum 17-OH progesterone and free testosterone levels in women patients with Familial Mediterranean Fever: a pivotal study.

Effects of Astragalus membranaceus roots supplementation on growth performance, serum antioxidant and immune response in finishing lambs

Polyunsaturated fatty acids augment tumoricidal action of 5-fluorouracil on gastric cancer cells by their action on vascular endothelial growth factor, tumor necrosis factor-α and lipid metabolism related factors

Therapeutic effects of ciprofloxacin/bushenhuazhuo combination on chronic prostatitis

Exhaled Hydrogen Sulfide Predicts Airway Inflammation Phenotype in COPD

Porcine parvovirus nonstructural protein NS1 activates NF-κB and it involves TLR2 signaling pathway

Evaluation of optimized continuous venovenous hemodiafiltration therapy efficiency in severe burn patients with sepsis

Hyperbaric oxygen on rehabilitation of brain tumors after surgery and effects on TNF-α and IL-6 levels

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22 Customer Q&As for Human TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

Question

Is EK0525 valid for human milk samples?

Verified customer

Asked: 2022-03-30

Answer

Human TNF Alpha/Tumor Necrosis Factor ELISA Kit PicoKine® (EK0525) has not been tested on human milk explicitly. In our experience customers have used it successfully in human milk. Thus we are willing to extend our quality guarantee to cover human milk as a viable sample type.

Boster Scientific Support

Answered: 2022-03-30

Question

The EK0525 manual indicates sensitivity as <1 pg/ml. Is it possible to see the signal difference at small concentrations using more sensitive detection reagent?

Verified customer

Asked: 2021-01-18

Answer

The sensitivity is the lower limit of target protein that can be detected by the kit. It is determined by adding two standard deviations to the mean O.D. value of twenty (20) blank wells and calculating the corresponding concentration. At the sensitivity level, the kit can detect the presence of the protein but not accurately quantify it. Accurate quantitation begins at the lowest point on the standard curve. For the Human TNF Alpha ELISA Kit PicoKine™ (EK0525), the values lower than 15.6pg/ml might not be reliable. If the user would like to see the signal difference at small concentrations, ELISA kits with higher sensitivity provided by other vendors are suggested to be used.

Boster Scientific Support

Answered: 2021-01-19

Question

Q: we need your suggestion regarding the dilution ratio of serum samples for detection of TNF-alpha in Human plasma? I am trying to measure a multiple analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2020-12-09

Answer

A: unable to understand the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the TNF-alpha ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2020-12-09

Question

Can EK0525 be used in calculating the tnf alpha concentration in human gingival cervicular fluid?

Verified customer

Asked: 2020-10-13

Answer

The Human TNF Alpha ELISA Kit PicoKine™ (EK0525) hasn't been tested with human gingival cervicular fluid.We suggest to run a pilot test for this. In theory, if the expression level of the target protein in the urine samples falls in the assay range of the kits, the kits are able to detect the protein in the sample.

Boster Scientific Support

Answered: 2020-10-14

Question

What dilution is needed for human serum samples?

Verified customer

Asked: 2020-09-25

Answer

For human serum samples using the Human TNF Alpha ELISA Kit PicoKine™ (EK0525), dilution of 1:1 is recommended.

Boster Scientific Support

Answered: 2020-09-29

Question

Does EK0525, EK0392, EK0373 react with BSA?

Verified Customer

Verified customer

Asked: 2020-04-21

Answer

EK0525, EK0392, EK0373 does not react with BSA.

Boster Scientific Support

Answered: 2020-04-21

Question

For EK0525, EK0392, EK0373, do these kits have binding for the forms of cytokine other than just the active form?

Verified Customer

Verified customer

Asked: 2020-01-15

Answer

EK0525, EK0392, EK0373 are able to detect total forms of TNF-Alpha, IL-1Beta, and IFN-Gamma.

Boster Scientific Support

Answered: 2020-01-15

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2020-01-06

Answer

A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2020-01-06

Question

What is the expiration date of EK0525 from the time of manufacturing, 6 months or 1 year?

Verified customer

Asked: 2019-12-17

Answer

Expiration is based on storage temperature. Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles.

Boster Scientific Support

Answered: 2019-12-17

Question

Can I calculate the results of unknown samples using a computer program with a 4-parameter logistic function? Keyword: data analysis

Verified Customer

Verified customer

Asked: 2019-12-16

Answer

Yes, 4-parameter logistic function can be used to calculate sample data.

Boster Scientific Support

Answered: 2019-12-16

Question

Will EK0525 work on Hep2G cell supernatant?

Verified customer

Asked: 2019-10-15

Answer

Yes, the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) would work with Hep2G cell supernatant.

Boster Scientific Support

Answered: 2019-10-16

Question

Will EK0525 work on Cell lysate samples?

Verified customer

Asked: 2019-10-15

Answer

The immunogen sequence of the antibodies in the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) is V77-L233 which is the soluble form of TNF alpha. The protein is synthesized in the intracellular space of cells and secretes into the extracellular space which is cell culture supernatant. And the kit might be able to detect partial intracellular secreted protein which has not released into extracellular space in cell lysate.

Boster Scientific Support

Answered: 2019-10-16

Question

Q: Can TNF-alpha ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2019-06-19

Answer

A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not suitable for other sample types than we have tested: it means further investigation is required. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please see the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.

Boster Scientific Support

Answered: 2019-06-19

Question

What is the assay range of EK0525 in cell culture supernates sample?

Verified customer

Asked: 2019-05-09

Answer

The assay range of the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) human TNF alpha kit in cell culture supernates is 15.6pg/ml-1000pg/ml.

Boster Scientific Support

Answered: 2019-05-10

Question

What is the assay range of EK0525 in serum and plasma samples?

Verified customer

Asked: 2019-05-09

Answer

The assay range of the Human TNF Alpha ELISA Kit PicoKine™ (EK0525) human TNF alpha kit in serum and plasma samples is 7.8pg/ml-500pg/ml.

Boster Scientific Support

Answered: 2019-05-10

Question

Q: how much samples can be assayed in a Picokine® ELISA Kit?

N. Phillips

Verified customer

Asked: 2019-04-22

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may might differ by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2019-04-22

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis TNF-alpha using the EK0525 Human TNF-alpha Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-03-14

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples in order to limit protein degradation.

Boster Scientific Support

Answered: 2019-03-14

Question

Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2017-07-31

Answer

A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2017-07-31

Question

Does EK0525 work with whole cell lysates?

Verified Customer

Verified customer

Asked: 2017-06-05

Answer

EK0525 is not suitable for working with whole cell lysates.

Boster Scientific Support

Answered: 2017-06-05

Question

Q: how to analyze ELISA data? I have obtained TNF-alpha level in plasma.

Verified Customer

Verified customer

Asked: 2017-01-13

Answer

A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2017-01-13

Question

For EK0525, a customer wanted to know if the kit would work on Cell lysate samples. They also wanted to know if the kit would work with Hep2G cell supernatant.

Verified Customer

Verified customer

Asked: 2014-09-04

Answer

The immunogen sequence of the antibodies in the kits is V77-L233 which is the soluble form of TNF alpha. The protein is synthesized in the intracellular space of cells and secretes into the extracellular space which is cell culture supernatant. And the kit might be able to detect patial intracellular secreted protein which hasn't released into extracellular space in cell lysate. The kit would work with Hep2G cell supernatant.

Boster Scientific Support

Answered: 2014-09-04

Question

I want to know more about your product EK0525 Human TNF Alpha ELISA Kit on the sensitivity of the assay. Do you have a detectable analytic range for urine samples? If not for urine then for serum?

Verified Customer

Verified customer

Asked: 2013-03-29

Answer

Our product EK0525 Human TNF Alpha ELISA Kit is not specifically validated with urine (only serum, plasma, and cell culture supernatants). However, we do not have concentration levels to report. When we tested human serum with EK0525 Human TNF Alpha ELISA Kit we found the measured levels were below the sensitivity limit which is 20 pg/ml not 1pg/ml as specified in the protocol. Thus, if testing body fluids, we would recommend using EK0525 Human TNF Alpha ELISA Kit mainly for spiking and recovery studies which have expected elevated TNF-alpha levels than normal.

Boster Scientific Support

Answered: 2013-03-29

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