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Product Info Summary
| SKU: | EK0920 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human TNFSF12/Tweak ELISA Kit PicoKine®
View all TWEAK/TNFSF12 ELISA kits
SKU/Catalog Number
EK0920
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human TNFSF12/Tweak ELISA Kit PicoKine™ (96 Tests). Quantitate Human TNFSF12 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/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 TNFSF12/Tweak ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0920)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: E.coli; Immunogen sequence: S94-H249
Sensitivity
<10 pg/ml
Assay Range
62.5 pg/ml - 4,000 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 EK0920 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0920 is reactive to TNFSF12 in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0920 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of TWEAK/TNFSF12
Tumor necrosis factor ligand superfamily member 12 also known as TNF-related weak inducer of apoptosis (TWEAK) is a protein that in humans is encoded by the TNFSF12 gene. The protein encoded by this gene is a cytokine that belongs to the tumor necrosis factor (TNF) ligand family. This protein is a ligand for the FN14/TWEAKR receptor. This cytokine has overlapping signaling functions with TNF, but displays a much wider tissue distribution. This cytokine can induce apoptosis via multiple pathways of cell death in a cell type-specific manner. This cytokine is also found to promote proliferation and migration of endothelial cells, and thus acts as a regulator ofangiogenesis. The TNFSF12 gene lies 878 bp upstream of the putative transcriptional start site of the TNFSF13 gene on chromosome 17p13.1.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0920-CAP | Anti-Human TNFSF12 Pre-coated 96-well strip microplate | 1 |
| EK0920-ST | Human TNFSF12 Standard | 2 vials, 10 ng/tube |
| EK0920-DA | Human TNFSF12 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 | 62.5 | 125 | 250 | 500 | 1000 | 2000 | 4000 | |
| O.D. | 0.02 | 0.088 | 0.151 | 0.26 | 0.507 | 0.881 | 1.401 | 2.174 |
Data Example Images
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Human TNFSF12 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect TWEAK/TNFSF12, Dilution ratio of 1:1, concentration in serum and plasma is 200-1300 pg/ml..
Some articles we found to cite concentrations of TWEAK/TNFSF12 in samples: 23800379 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human TNFSF12/Tweak 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) | 160 | 584 | 1794 | 172 | 587 | 1626 |
| Standard deviation | 9.12 | 28.61 | 91.49 | 12.55 | 36.98 | 87.8 |
| CV (%) | 5.7% | 4.9% | 5.4%% | 7.3% | 6.3% | 5.4% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of TWEAK/TNFSF12 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 | 160 | 163 | 177 | 179 | 169 | 8.34 | 4.9% |
| Sample 2 | 584 | 541 | 582 | 513 | 555 | 29.7 | 5.3% |
| Sample 3 | 1794 | 2011 | 2021 | 1798 | 1906 | 110.06 | 5.7% |
Protein Target Info & Infographic
Gene/Protein Information For TNFSF12 (Source: Uniprot.Org, NCBI)
Gene Name
TNFSF12
Full Name
Tumor necrosis factor ligand superfamily member 12
Weight
27.216kDa
Superfamily
tumor necrosis factor family
Alternative Names
APO3 ligand; APO3LMGC20669; DR3LGTWEAKAPO3/DR3 ligand; MGC129581; TNF-related weak inducer of apoptosis; TNFSF12; tumor necrosis factor (ligand) superfamily, member 12; tumor necrosis factor ligand superfamily member 12; TWEAK TNFSF12 APO3L, DR3LG, TNLG4A, TWEAK TNF superfamily member 12 tumor necrosis factor ligand superfamily member 12|APO3 ligand|APO3/DR3 ligand|TNF-related WEAK inducer of apoptosis|tumor necrosis factor (ligand) superfamily, member 12|tumor necrosis factor ligand 4A|tumor necrosis factor superfamily member 12
*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 TNFSF12, check out the TNFSF12 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 TNFSF12: 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 Human TNFSF12/Tweak ELISA Kit PicoKine® (EK0920)
Hello CJ!
EK0920 has been cited in 1 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.
How much apoptosis does carbon monoxide poisoning cause? Primary clinical soluble TWEAK protein level study:
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Customer Reviews
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Customer Q&As
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Find answers in Q&As, reviews.
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Submit your question
11 Customer Q&As for Human TNFSF12/Tweak ELISA Kit PicoKine®
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
C. Bhatt
Verified customer
Asked: 2020-03-20
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: 2020-03-20
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of TWEAK in Human serum? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2019-11-15
Answer
A: unable to know 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 TWEAK 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: 2019-11-15
Question
Dear Support: Do you have any idea if this EK0920 antibody would also react with murine Fn14 or it is only specific to human Fn14 isoforms? If it is specific to human Fn14 with no cross-reactivity with murine Fn14 what would you recommend for an antibody that can be used to neutralize the murine Fn14 cell lines?
Verified Customer
Verified customer
Asked: 2019-08-22
Answer
Our product EK0920 has an immunogen sequence of E.coli expressed recombinant protein with a sequence MKGRKTRARR AIAAHYEVHP RPGQDGAQAG VDGTVSGWEE ARINSSSPLR YNRQIGEFIV TRAGLYYLYC QVHFDEGKAV YLKLDLLVDG VLALRCLEEF SATAASSLGP QLRLCQVSGL LALRPGSSLR IRTLPWAHLK AAPFLTYFGL FQVH. The sequence is 92% similar with mouse sequence, therefore EK0920 will most probably cross react with the mouse species. We can guarantee that this EK0920 antibody has been successfully tested in functional assays on human cells.
Boster Scientific Support
Answered: 2019-08-22
Question
Q: how to analyze ELISA data? I have obtained TWEAK level in serum.
A. Clark
Verified customer
Asked: 2019-07-25
Answer
A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2019-07-25
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-07-01
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 vary slightly by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2019-07-01
Question
Q: if the enzyme conjugated TWEAK 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 TWEAK antigen?
Verified Customer
Verified customer
Asked: 2018-10-24
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: 2018-10-24
Question
Q: Can TWEAK ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2018-08-26
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: 2018-08-26
Question
Q: What is the optimal O.D. value for TWEAK ELISA kit? I performed your TWEAK 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 TWEAK even though the O.D. values are not very high?
K. Turner
Verified customer
Asked: 2018-07-01
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. a point of focus should be 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 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 changeing 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 TWEAK ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of TWEAK that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2018-07-01
Question
Q: is it okay to use citrate plasma as samples in Human TWEAK Picokine® ELISA Kit (Catalog # EK0920)?
V. Puri
Verified customer
Asked: 2016-07-01
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of TWEAK causing disruption of its protein structure. TWEAK may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the TWEAK ELISA, treating samples with a number of 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: 2016-07-01
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2016-04-20
Answer
A: in 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: 2016-04-20
Question
Q: which procedure should I follow in order to thaw whole blood sample for TWEAK ELISA after freezing?
Verified Customer
Verified customer
Asked: 2016-04-08
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 TWEAK 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: 2016-04-08
