Human TNFRSF25/DR3 ELISA Kit PicoKine®

DR3/TNFRSF25 ELISA kit for Human

Human TNFRSF25/DR3 ELISA Kit PicoKine™ (96 Tests). Quantitate Human TNFRSF25 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1579
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 Name

Human TNFRSF25/DR3 ELISA Kit PicoKine®

View all DR3/TNFRSF25 ELISA kits

SKU/Catalog Number

EK1579

Size

96 wells/kit, with removable strips.

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

Description

Human TNFRSF25/DR3 ELISA Kit PicoKine™ (96 Tests). Quantitate Human TNFRSF25 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 TNFRSF25/DR3 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1579)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: Q25-F201

Sensitivity

<10 pg/ml

Assay Range

312 pg/ml - 20,000 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 EK1579 for more details

Cross-reactivity

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

Reactive Species

EK1579 is reactive to TNFRSF25 in Human samples

Validated Sample Types

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

Application Guarantee

EK1579 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of DR3/TNFRSF25

Death receptor 3 (DR3), also known as tumor necrosis factor receptor superfamily member 25 (TNFRSF25), is a cell surface receptor of the tumor necrosis factor receptor superfamily which mediates apoptotic signalling and differentiation. Knockout studies in mice suggested the role of this gene in the removal of self-reactive T cells in the thymus. Multiple alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported, most of which are potentially secreted molecules. The alternative splicing of this gene in B and T cells encounters a programmed change upon T-cell activation, which predominantly produces full-length, membrane bound isoforms, and is thought to be involved in controlling lymphocyte proliferation induced by T-cell activation.

Kit Components

Catalog Number Description Quantity
EK1579-CAP Anti-Human TNFRSF25 Pre-coated 96-well strip microplate 1
EK1579-ST Human TNFRSF25 Standard 2 vials, 20 ng/tube
EK1579-DA Human TNFRSF25 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)03126251250250050001000020000
O.D.0.0470.1170.1480.3360.4290.7241.2552.087

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect DR3/TNFRSF25, Dilution ratio of 1:1, concentration in serum and plasma is less than the lowest standard, 312 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human TNFRSF25/DR3 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)69124879783668243010603
Standard deviation37.31154.19489.1542.75196.83689.19
CV (%)5.4%6.2%5%6.4%8.1%6.5%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of DR3/TNFRSF25 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 169163963966965921.923.3%
Sample 22487231022532294233689.623.8%
Sample 39783100469881101039953127.71.2%
*number of samples for each test n=16.

Gene/Protein Information For TNFRSF25 (Source: Uniprot.Org, NCBI)

Gene Name

TNFRSF25

Full Name

Tumor necrosis factor receptor superfamily member 25

Weight

45.385kDa

Alternative Names

APO3; Apo-3; Apoptosis-inducing receptor AIR; Apoptosis-mediating receptor DR3; Apoptosis-mediating receptor TRAMP; Death receptor 3; DR3; DR3member 12 (translocatingchain-association membrane protein); LARD; Lymphocyte-associated receptor of death; Protein WSL; Protein WSL-1; TNFRSF25; TRAMP; tumor necrosis factor receptor superfamily, member 25; WSL; WSL1; WSL-1 TNFRSF25 APO-3, DDR3, DR3, GEF720, LARD, PLEKHG5, TNFRSF12, TR3, TRAMP, WSL-1, WSL-LR TNF receptor superfamily member 25 tumor necrosis factor receptor superfamily member 25|Guanine nucleotide exchange factor 720|PH domain-containing family G member 5|Pleckstrin homology domain-containing family G member 5|apoptosis inducing receptor|apoptosis-inducing receptor AIR|apoptosis-mediating receptor DR3|apoptosis-mediating receptor TRAMP|death domain receptor 3 soluble form|death receptor beta|lymphocyte-associated receptor of death|protein WSL-1|tumor necrosis factor receptor superfamily, member 12 (translocating chain-association membrane protein)

*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 TNFRSF25, check out the TNFRSF25 Infographic

TNFRSF25 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 TNFRSF25: 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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8 Customer Q&As for Human TNFRSF25/DR3 ELISA Kit PicoKine®

Question

Q: What is the optimal O.D. value for DR3 ELISA kit? I performed your DR3 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 DR3 even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2020-04-29

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 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 DR3 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of DR3 that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2020-04-29

Question

Q: can you suggest the dilution ratio of serum samples for detection of DR3 in Human serum? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

W. Jones

Verified customer

Asked: 2019-04-13

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 DR3 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: 2019-04-13

Question

Q: Can DR3 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2019-01-04

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: 2019-01-04

Question

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

L. Edwards

Verified customer

Asked: 2018-12-10

Answer

A: in 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: 2018-12-10

Question

Q: if the enzyme conjugated DR3 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 DR3 antigen?

G. Dhar

Verified customer

Asked: 2018-07-15

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: 2018-07-15

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification DR3 using the EK1579 Human DR3 Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2017-07-14

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may alter 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. avoid repeat freeze-thaw cycles with the stored samples to limit protein degradation.

Boster Scientific Support

Answered: 2017-07-14

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2017-01-12

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: 2017-01-12

Question

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

Verified Customer

Verified customer

Asked: 2015-09-12

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: 2015-09-12

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