Mouse CD40/TNFRSF5 ELISA Kit PicoKine®

CD40/TNFRSF5 ELISA kit for Mouse

Mouse CD40/TNFRSF5 ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse Cd40 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 1pg/ml.

Product Info Summary

SKU: EK0703
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 CD40/TNFRSF5 ELISA Kit PicoKine®

View all CD40/TNFRSF5 ELISA kits

SKU/Catalog Number

EK0703

Size

96 wells/kit, with removable strips.

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

Description

Mouse CD40/TNFRSF5 ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse Cd40 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 CD40/TNFRSF5 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0703)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: V24-R193

Sensitivity

<1 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 EK0703 for more details

Cross-reactivity

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

Reactive Species

EK0703 is reactive to CD40 in Mouse samples

Validated Sample Types

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

Application Guarantee

EK0703 is guaranteed for ELISA in Mouse by Boster Guarantee

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

Background of CD40/TNFRSF5

CD40 is a cell surface receptor that is expressed on the surface of all mature B cells, most mature B-cell malignancies, and some early B-cell acute lymphocytic leukemias, but is not expressed on plasma cells. The CD40 gene is localized to the long arm of chromosome 20, bands q12-q13.2. The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation.

Kit Components

Catalog Number Description Quantity
EK0703-CAP Anti-Mouse CD40 Pre-coated 96-well strip microplate 1
EK0703-ST Mouse CD40 Standard 2 vials, 10 ng/tube
EK0703-DA Mouse CD40 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.0.0040.0690.1350.2300.4950.9051.4212.050

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect CD40/TNFRSF5, Dilution ratio of 1:1, concentration in serum and plasma is 39-183g/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Mouse CD40/TNFRSF5 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)3020842127195393
Standard deviation1.8610.8118.11.8612.0922.79
CV (%)6.2%5.2%4.3%6.9%6.2%5.8%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of CD40/TNFRSF5 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 130293129290.822.8%
Sample 22081922002072016.413.1%
Sample 342144450847946333.187.1%
*number of samples for each test n=16.

Gene/Protein Information For CD40 (Source: Uniprot.Org, NCBI)

Gene Name

CD40

Full Name

Tumor necrosis factor receptor superfamily member 5

Weight

30.619kDa

Alternative Names

B cell surface antigen CD40; B-cell surface antigen CD40; Bp50B cell-associated molecule; CD40 antigen; CD40 molecule, TNF receptor superfamily member 5; CD40 type II isoform; CD40; CD40L receptor; CDw40; MGC9013; nerve growth factor receptor-related B-lymphocyte activation molecule; p50; TNFRSF5; TNFRSF5CD40 antigen (TNF receptor superfamily member 5); tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5 CD40 Bp50, CDW40, TNFRSF5, p50 CD40 molecule tumor necrosis factor receptor superfamily member 5|B cell surface antigen CD40|B cell-associated molecule|CD40 molecule, TNF receptor superfamily member 5|CD40L receptor

*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 CD40, check out the CD40 Infographic

CD40 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 CD40: 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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13 Customer Q&As for Mouse CD40/TNFRSF5 ELISA Kit PicoKine®

Question

Q: Can CD40 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Y. Miller

Verified customer

Asked: 2020-12-07

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 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: 2020-12-07

Question

Q: What is the optimal O.D. value for CD40 ELISA kit? I used your CD40 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 CD40 even though the O.D. values are not very high?

P. Allen

Verified customer

Asked: 2020-08-20

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 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 CD40 ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of CD40 that can be declared/interpreted as positive by the above standard is 1pg/ml.

Boster Scientific Support

Answered: 2020-08-20

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2020-06-27

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: 2020-06-27

Question

Q: how do I analyze ELISA data? I have obtained CD40 level in cell lysates.

Verified Customer

Verified customer

Asked: 2019-12-23

Answer

A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2019-12-23

Question

Could you please tell me what was done to validate that this antibody clone Human CD40/TNFRSF5 ELISA Kit is really directed against CD40.

Verified Customer

Verified customer

Asked: 2019-12-17

Answer

Human CD40/TNFRSF5 to CD40 has been characterized as part of the Leucocyte Workshop VI and the testing was done via IHC (frozen sections), Western blot and functional studies. However, for your further references, we listed several published papers about this antibody. One of the reference discuss about the original characterization of this clone, do check this; Vall A et al. Activation of human B lymphocytes through CD40 and interleukin 4. Eur J Immunol 19:1463-7 (1989). PubMed: 2476318.

Boster Scientific Support

Answered: 2019-12-17

Question

I already received the Human CD40/TNFRSF5 ELISA Kit. I just want to know if the solution is sterile. Kind regards.

Verified Customer

Verified customer

Asked: 2019-08-22

Answer

Yes, Human CD40/TNFRSF5 ELISA Kit is sterile when we dispatched it to our customers. It is fully validated and guaranteed to use in Flow cytometry, functional studies and cellular activation

Boster Scientific Support

Answered: 2019-08-22

Question

Q: can you recommend the dilution ratio of serum samples for detection of CD40 in Mouse cell lysates? 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.

D. Krishna

Verified customer

Asked: 2019-01-24

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 CD40 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: 2019-01-24

Question

Q: how much samples can be assayed in a Picokine® ELISA Kit?

Z. Varadkar

Verified customer

Asked: 2018-09-25

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 might differ by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-09-25

Question

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

Verified Customer

Verified customer

Asked: 2018-02-23

Answer

A: for 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 can effect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2018-02-23

Question

Q: is it okay to use citrate plasma as samples in Mouse CD40 Picokine® ELISA Kit (Catalog # EK0703)?

Verified Customer

Verified customer

Asked: 2017-10-16

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of CD40 causing disruption of its protein structure. CD40 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 CD40 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: 2017-10-16

Question

Q: if the enzyme conjugated CD40 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 CD40 antigen?

Verified Customer

Verified customer

Asked: 2016-06-18

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: 2016-06-18

Question

I want to purchase the Mouse CD40/TNFRSF5 ELISA Kit to test it in serum samples. However the protocol says that I have to dilute the sample but it doesn't indicate which dilution exactly or a range. Can I have more information about it?

Verified Customer

Verified customer

Asked: 2016-04-25

Answer

We have not specific range for Mouse CD40/TNFRSF5 ELISA Kit, since this can vary significantly depending on the sample. We suggest to try multiple dilutions in a trial run to determine which dilution will be most suitable for your samples.

Boster Scientific Support

Answered: 2016-04-25

Question

For clarification; I would like to know if the Human CD40/TNFRSF5 ELISA Kit is for the detection of soluble CD40L?

Verified Customer

Verified customer

Asked: 2013-10-11

Answer

The Human CD40/TNFRSF5 ELISA Kit recognizes the extracellular region of CD40L (aa 47

Boster Scientific Support

Answered: 2013-10-11

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