Rat TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

TNF-alpha ELISA kit for Rat

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

Product Info Summary

SKU: EK0526
Size: 96 wells/kit, with removable strips.
Reactive Species: Rat
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

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Product Name

Rat TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

View all TNF-alpha ELISA kits

SKU/Catalog Number

EK0526

Size

96 wells/kit, with removable strips.

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

Description

Rat TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Tnf in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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

Rat TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0526)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: L80-L235

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

EK0526 is reactive to Tnf in Rat samples

Validated Sample Types

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

Application Guarantee

EK0526 is guaranteed for ELISA in Rat 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. The standard product used in this kit is recombinant rat TNFalpha, consisting of 157 amino acids with the molecular mass of 17KDa.

Kit Components

Catalog Number Description Quantity
EK0526-CAP Anti-Rat Tnf Pre-coated 96-well strip microplate 1
EK0526-ST Rat Tnf Standard 2 vials, 10 ng/tube
EK0526-DA Rat 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.0.0100.0310.0620.1320.2030.5550.8841.838

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 target in serum is below detection limit..

Some articles we found to cite concentrations of TNF-alpha in samples: 24744897 (Pubmed IDs).

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Rat 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)14962801398306
Standard deviation0.577.3920.440.787.5424.78
CV (%)4.1%7.7%7.3%6%7.7%8.1%

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 1141414141400%
Sample 2969392101953.53.6%
Sample 328025827430627917.286.1%
*number of samples for each test n=16.

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

Uniprot ID

P16599

Gene ID

24835

Gene Name

Tnf

Full Name

Tumor necrosis factor

Weight

25.806kDa

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|RATTNF, TNF-alphaa, Tnf|tumor necrosis factor|tumor necrosis factor|cachectin|tumor necrosis factor (TNF superfamily, member 2)|tumor necrosis factor ligand superfamily member 2

*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]

EK0526 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.

Promotion of chemerin in rat diabetic kidney disease through enhancement of TGF-β1/Smads/CTGF pathway

Salvianolic acid A attenuates early brain injury after subarachnoid hemorrhage in rats by regulating ERK/P38/Nrf2 signaling

Pachymic acid ameliorates sepsis-induced acute kidney injury by suppressing inflammation and activating the Nrf2/HO-1 pathway in rats.

Effect of early fluid resuscitation on the lung in a rat model of lipopolysaccharide-induced septic shock

Comparison of the Effects of Curcumin, Tramadol and Surgical Treatments on Neuropathic Pain Induced by Chronic Constriction Injury in Rats.

Characterization of astrocytes and microglial cells in the hippocampal CA1 region after transient focal cerebral ischemia in rats treated with Ilexonin A

Sublytic C5b-9 Induces Glomerular Mesangial Cell Apoptosis through the Cascade Pathway of MEKK2–p38 MAPK–IRF-1–TRADD–Caspase 8 in Rat Thy-1 Nephritis

Cyanidin‑3‑O‑β‑glucoside protects against pulmonary artery hypertension induced by monocrotaline via the TGF‑β1/p38 MAPK/CREB signaling pathway

Tanshinone IIA attenuates cerebral aneurysm formation by inhibiting the NF‑κB‑mediated inflammatory response

Protective effects of propofol on experimental neonatal acute lung injury

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11 Customer Q&As for Rat TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®

Question

Does EK0526 contain any recombinant proteins expressed in baculoviral system? Which kit components are recombinant proteins and what species?

Verified customer

Asked: 2021-11-25

Answer

The standard in the Rat TNF Alpha ELISA Kit PicoKine™ (EK0526) is a recombinant rat protein expressed in E.coli system.

Boster Scientific Support

Answered: 2021-12-01

Question

Q: What is the optimal O.D. value for TNF-alpha ELISA kit? I performed your TNF-alpha ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. is it safe to say these samples contain TNF-alpha even though the O.D. values are not very high?

L. Anand

Verified customer

Asked: 2020-12-21

Answer

A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. what you should focus on is whether your sample O.D. values are statistically significantly higher than your blank values. in the above example, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this TNF-alpha ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of TNF-alpha that can be declared/interpreted as positive by the above standard is 1pg/ml.

Boster Scientific Support

Answered: 2020-12-21

Question

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

Verified Customer

Verified customer

Asked: 2020-01-11

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 assessd 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 view 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: 2020-01-11

Question

Q: is it okay to use citrate plasma as samples in Rat TNF-alpha Picokine® ELISA Kit (Catalog # EK0526)?

Verified Customer

Verified customer

Asked: 2019-11-23

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of TNF-alpha causing degradation of its protein structure. TNF-alpha may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the TNF-alpha ELISA, treating samples with different anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2019-11-23

Question

Is EK0526 able to detect the non-soluble form of TNF alpha?

Verified Customer

Verified customer

Asked: 2019-10-11

Answer

EK0526 is able to detect the non-soluble form of TNF alpha.

Boster Scientific Support

Answered: 2019-10-11

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

G. Moore

Verified customer

Asked: 2019-08-23

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 required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2019-08-23

Question

To Boster Support: I have a question about EK0526 Rat TNF Alpha ELISA Kit, what is the minimum detectable concentration of TNF alpha? I want to be able to detect as little as 5 pg/ml.

Verified Customer

Verified customer

Asked: 2019-08-22

Answer

The limit of detection of EK0526 Rat TNF Alpha ELISA Kit is 15 pg/ml, which is reached at by taking the mean optical density of 64 blank wells (no TNF alpha), then add two standard deviations of the mean, then calculating the corresponding concentration from the standard curve, including one or more standard dilutions will not improve the sensitivity.

Boster Scientific Support

Answered: 2019-08-22

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification TNF-alpha using the EK0526 Rat TNF-alpha Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-01-27

Answer

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

Boster Scientific Support

Answered: 2019-01-27

Question

Q: how to thaw whole blood sample for TNF-alpha ELISA after freezing?

R. Wang

Verified customer

Asked: 2018-10-10

Answer

A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test TNF-alpha for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

Boster Scientific Support

Answered: 2018-10-10

Question

Q: we need your suggestion regarding the dilution ratio of serum samples for detection of TNF-alpha in Rat cell lysates? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.

C. Collins

Verified customer

Asked: 2018-09-16

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: 2018-09-16

Question

I want to measure Rat TNF Alpha using your product EK0526 Rat TNF Alpha ELISA Kit in rat hippocampus tissues. I read the manual and it stated that the EK0526 Rat TNF Alpha ELISA Kit can measure these cytokines level but it is not specified in what kind of buffer it must be homogenize the tissues. My concern is that; Can I use using homgenates in Lysis buffer 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0)?

Verified Customer

Verified customer

Asked: 2016-10-13

Answer

That lysis buffer is good, the 150 mM sodium chloride, 1.0% Triton X-100, 50 mM Tris pH 8.0. Please check these general rules: Do not use >0.1% SDS or other strongly denaturing detergents. Non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will also work. Use no more than 2% v/v total detergent. Do not use sodium azide and avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols. Hence, e strongly recommend to add a protease inhibitor cocktail to the lysis buffer prior to homogenization.

Boster Scientific Support

Answered: 2016-10-13

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