Mouse TNFSF12/Tweak ELISA Kit PicoKine®

TWEAK/TNFSF12 ELISA kit for Mouse

Mouse TNFSF12/Tweak ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse Tnfsf12 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1176
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 Name

Mouse TNFSF12/Tweak ELISA Kit PicoKine®

View all TWEAK/TNFSF12 ELISA kits

SKU/Catalog Number

EK1176

Size

96 wells/kit, with removable strips.

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

Description

Mouse TNFSF12/Tweak ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse 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

Mouse TNFSF12/Tweak ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1176)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: S94-H249

Sensitivity

<10 pg/ml

Assay Range

15.6 pg/ml - 1,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

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 EK1176 for more details

Cross-reactivity

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

Reactive Species

EK1176 is reactive to TNFSF12 in Mouse samples

Validated Sample Types

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

Application Guarantee

EK1176 is guaranteed for ELISA in Mouse 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
EK1176-CAP Anti-Mouse TNFSF12 Pre-coated 96-well strip microplate 1
EK1176-ST Mouse TNFSF12 Standard 2 vials, 10 ng/tube
EK1176-DA Mouse 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.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015.631.262.51252505001000
O.D.0.0250.1010.1900.3160.5571.0181.7512.543

Data Example Images

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 around 70 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Mouse 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 PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)3919243342205410
Standard deviation1.839.7918.612.4710.4522.14
CV (%)4.7%5.4%4.3%5.9%5.4%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.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 139404237391.84.6%
Sample 21922121921991988.164.1%
Sample 343349449450148027.575.7%
*number of samples for each test n=16.

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

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].

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10 Customer Q&As for Mouse TNFSF12/Tweak ELISA Kit PicoKine®

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

Verified Customer

Verified customer

Asked: 2020-10-31

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: 2020-10-31

Question

Q: how can I thaw whole blood sample for TWEAK ELISA after freezing?

Verified Customer

Verified customer

Asked: 2020-10-26

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 separate the serum to freeze for later analysis.

Boster Scientific Support

Answered: 2020-10-26

Question

Can serum samples be diluted using EK1176?

Verified customer

Asked: 2020-09-08

Answer

The suggested dilution ratio for serum is 1:1 while using the Mouse TNFSF12/Tweak ELISA Kit PicoKine™ (EK1176).

Boster Scientific Support

Answered: 2020-09-11

Question

Q: how to analyze ELISA data? I measured TWEAK level in plasma.

S. Hall

Verified customer

Asked: 2020-03-23

Answer

A: we recommend you 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: 2020-03-23

Question

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

Verified Customer

Verified customer

Asked: 2019-11-17

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 needed. 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 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: 2019-11-17

Question

Q: What is the optimal O.D. value for TWEAK ELISA kit? I used 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?

Verified Customer

Verified customer

Asked: 2019-09-20

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. in your 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 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 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: 2019-09-20

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification TWEAK using the EK1176 Mouse TWEAK Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-04-16

Answer

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

Boster Scientific Support

Answered: 2019-04-16

Question

Q: can you suggest the dilution ratio of serum samples for detection of TWEAK in Mouse plasma? I am trying to measure a a number of 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.

L. Lewis

Verified customer

Asked: 2018-02-24

Answer

A: having little idea about 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: 2018-02-24

Question

Q: if the enzyme conjugated TWEAK antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the TWEAK antigen?

S. Mangal

Verified customer

Asked: 2017-02-20

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: 2017-02-20

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

B. Babu

Verified customer

Asked: 2014-09-21

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 may affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2014-09-21

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