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Product Info Summary
| SKU: | EK0527 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Mouse |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®
SKU/Catalog Number
EK0527
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse 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
Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0527)
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
Standard Dilution Instructions
Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK0527 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0527 is reactive to TNF in Mouse samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0527 is guaranteed for ELISA in Mouse 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 |
|---|---|---|
| EK0527-CAP | Anti-Mouse TNF Pre-coated 96-well strip microplate | 1 |
| EK0527-ST | Mouse TNF Standard | 2 vials, 10 ng/tube |
| EK0527-DA | Mouse 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.
- Incubator.
- 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.
Innovating Scientists Reward
If you are the first to review this product, or if you have results for a special sample, species or application this product is not validated in, share your results with us and receive product credits you can use towards any Boster products! Applicable to all scientists world wide.
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Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.051 | 0.090 | 0.156 | 0.281 | 0.443 | 0.723 | 1.284 | 1.958 |
Data Example Images
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Mouse TNF alpha PicoKine ELISA Kit standard curve
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 and plasma is less than the lowest standard, 15.6 pg/ml..
Some articles we found to cite concentrations of TNF-alpha in samples: 25561799 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Mouse 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 Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 18 | 50 | 299 | 16 | 53 | 305 |
| Standard deviation | 0.82 | 3.9 | 22.12 | 0.91 | 4.39 | 24.4 |
| CV (%) | 4.6% | 7.8% | 7.4% | 5.7% | 8.3% | 8% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of TNF-alpha in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 18 | 18 | 17 | 19 | 18 | 0.7 | 3.8% |
| Sample 2 | 50 | 54 | 54 | 47 | 51 | 2.94 | 5.7% |
| Sample 3 | 299 | 294 | 293 | 285 | 292 | 5.01 | 1.7% |
Protein Target Info & Infographic
Gene/Protein Information For TNF (Source: Uniprot.Org, NCBI)
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
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].
Specific Publications For Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine® (EK0527)
Hello CJ!
EK0527 has been cited in 894 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.
Propofol alleviates hepatic ischemia/reperfusion injury via the activation of the Sirt1 pathway
Catechin Weakens Diabetic Retinopathy by Inhibiting the Expression of NF-κB Signaling Pathway-Mediated Inflammatory Factors
Luteolin alleviates NLRP3 inflammasome activation and directs macrophage polarization in lipopolysaccharide-stimulated RAW264.7 cells
Increased expression of CXCR5 and CXCL13 in mice with experimental autoimmune myocarditis.
IL-6 induced lncRNA MALAT1 enhances TNF-α expression in LPS-induced septic cardiomyocytes via activation of SAA3.
MicroRNA-155 in exosomes secreted from helicobacter pylori infection macrophages immunomodulates inflammatory response
Cold exposure stimulates lipid metabolism, induces inflammatory response in the adipose tissue of mice and promotes the osteogenic differentiation of BMMSCs via the p38 MAPK pathway in vitro
Unfractionated heparin attenuates intestinal injury in mouse model of sepsis by inhibiting heparanase
The role of RIP1 and RIP3 in the development of aplastic anemia induced by cyclophosphamide and busulphan in mice
Toll-like receptor 4 implicated in acute lung injury induced by paraquat poisoning in mice
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Customer Reviews
Have you used Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®?
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3 Reviews For Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®
0
Great TNF-Alpha Kit from Boster!--Harrison Esterly, Appalachian State University, Chemistry, Lab Technician
Excellent
Source: Biocompare.com
| Application | ELISA |
|---|---|
| Sample | Mouse serum |
| Detection | -- |
"The kit contained an extra standard which is very useful if you need to optimize your dilution factor before you use the rest of your samples. The instructions were very easy to follow and the results we obtained were consistent between replicates. Very easy to use. Don't waste your time or money [on other kits]; just use this kit." (Customer’s Tip: "Optimization dilution factors should range from 1:5 to 1:200.")
Harrison Esterly
Verified customer
Submitted 2019-12-06
0
Excellent Quantification In Vitro and In Vivo--Divya Sharma, NDSU, Pharmaceutical Sciences, Graduate Student
Excellent
Source: Biocompare.com
| Application | ELISA |
|---|---|
| Sample | Cell culture media, mouse serum |
| Detection | -- |
"I was able to estimate TNFa in cell culture media before and after treatment with lipopolysaccharides in primary macrophage cell line. In vivo estimation was done in control and treated group of mice (high fat diet treatment) by collecting blood, separating serum, and estimating the cytokine levels in serum. Overall reliable estimation was achieved. Reliable reproducible result and brand of ELISA kits. Good ELISA kit. Reliable vendor. Good continuity between different batches of ELISA kits. Will continue to buy from Boster Biological." (Customer’s Tip: "Do a test run of sample and quantities may be higher than the standard plot range.")
Divya Sharma
Verified customer
Submitted 2019-05-21
0
Great TNF alpha Kit--Tho Pham, UCONN, NUTRITIONAL SCIENCES, GA
Excellent
Source: Biocompare.com
| Applications | ELISA |
|---|---|
| Sample | Cell culture supernatant |
| Detection | -- |
"This ELISA kit worked as advertised. Wells can be removed as strips, so not everything has to be used. Great product. Results are good and have low %CV. Quick assay kit and cheaper than other companies in my opinon."
Tho Pham
Verified customer
Submitted 2015-12-29
Customer Q&As
Have a question?
Find answers in Q&As, reviews.
Can't find your answer?
Submit your question
14 Customer Q&As for Mouse TNF Alpha/Tumor necrosis factor ELISA Kit PicoKine®
Question
What host species of antibody used in EK0527- TNFa (mouse) ELISA Kit?
Verified customer
Asked: 2020-06-18
Answer
For the Mouse TNF Alpha ELISA Kit PicoKine™ (EK0527), Host of capture antibody: Hamster Host of detection antibody: Goat
Boster Scientific Support
Answered: 2020-06-18
Question
Q: if the enzyme conjugated TNF-alpha antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the TNF-alpha antigen?
Y. Banerjee
Verified customer
Asked: 2020-05-09
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2020-05-09
Question
Q: can I use citrate plasma as samples in Mouse TNF-alpha Picokine® ELISA Kit (Catalog # EK0527)?
W. Gonzalez
Verified customer
Asked: 2019-12-03
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can attach 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 needed for detection and block the antigen antibody reaction. We have tested the TNF-alpha ELISA, treating samples with various 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: 2019-12-03
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2019-11-21
Answer
A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions
Boster Scientific Support
Answered: 2019-11-21
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. can I consider these samples contain TNF-alpha even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-09-26
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. regarding your protocol, 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 proportion to your positive controls. normally, 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: 2019-09-26
Question
Can you confirm if EK0394,EK0568,EK0527,EK0411 and EK0596 are compatible with ELISA microplate washers?
Verified customer
Asked: 2019-05-30
Answer
Our ELISA kits should be compatible for all ELISA microplate washers as long as it supports a 96 well plate.
Boster Scientific Support
Answered: 2019-05-30
Question
Can you confirm if the EK0527 is compatible with ELISA microplate washers?
Verified customer
Asked: 2019-05-30
Answer
Our ELISA kits should be compatible for all ELISA microplate washers as long as it supports a 96 well plate.
Boster Scientific Support
Answered: 2019-05-30
Question
Can the sample diluent buffer, ABC diluent buffer and antibody diluent buffer be used interchangeably between the kit?
Verified customer
Asked: 2019-05-28
Answer
For the Mouse IL-1 Beta ELISA Kit PicoKine™ (EK0394), Mouse MCP-1 ELISA Kit PicoKine™ (EK0568), Mouse TNF Alpha ELISA Kit PicoKine™ (EK0527), Mouse IL-6 ELISA Kit PicoKine™ (EK0411) and Mouse Adiponectin ELISA Kit PicoKine™ (EK0596), the sample diluent buffer, ABC diluent buffer and antibody diluent buffer can be used interchangeably between the kit. However, the sample diluent buffer can only be used to dilute standards and samples, ABC diluent buffer is used to dilute ABC only and antibody diluent buffer is used to dilute antibody only.
Boster Scientific Support
Answered: 2019-05-29
Question
Is it possible for EK0527 to detect 0.5pg/ml concentrations of Mouse TNF Alpha? Do you have recommendations for a more sensitive kit?
Verified customer
Asked: 2019-05-06
Answer
Unfortunately, it would be difficult for the Mouse TNF Alpha ELISA Kit PicoKine™ (EK0527) to detect that concentration and we are unable to offer a more sensitive kit.
Boster Scientific Support
Answered: 2019-05-07
Question
Q: Can TNF-alpha ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2019-02-06
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 valid for other sample types than we have tested: it means further investigation is a must. 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 check 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: 2019-02-06
Question
I would like to ask at what concentration of Mouse TNF alpha is measured in normal mouse plasma samples? As I research in literature the values indicates 10-40 pg/mL but your product EK0527 Mouse TNF Alpha ELISA Kit shows <1 pg/mL. Can this still detect my samples? As to not waste any of my plasma.
Verified Customer
Verified customer
Asked: 2019-01-24
Answer
Yes, it can detect your samples and we would recommend the starting dilution factor is 2-fold to reduce the risk of matrix effects. So, there is a wide range of possible detected levels, and some troubleshooting and optimization may be necessary if the samples contain a low level of the analytes.
Boster Scientific Support
Answered: 2019-01-24
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
H. Patel
Verified customer
Asked: 2018-06-27
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 depends upon the kit used so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2018-06-27
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2018-03-06
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are included in the product insert. Components in lysate and lysis buffer can have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-03-06
Question
Q: which procedure should I follow in order to thaw whole blood sample for TNF-alpha ELISA after freezing?
Verified Customer
Verified customer
Asked: 2017-11-06
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 separate the serum to freeze for later analysis.
Boster Scientific Support
Answered: 2017-11-06
