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Product Info Summary
| SKU: | EK0573 |
|---|---|
| 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 CD40 Ligand/TNFSF5/CD40LG ELISA Kit PicoKine®
View all CD40 Ligand/TNFSF5 ELISA kits
SKU/Catalog Number
EK0573
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human CD40 Ligand/TNFSF5/CD40LG ELISA Kit PicoKine® (96 Tests). Quantitate Human CD40LG in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 15pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
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 CD40 Ligand/TNFSF5/CD40LG ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0573)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: E108-L261
Sensitivity
<15 pg/ml
Assay Range
62.5 pg/ml - 4,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0573 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0573 is reactive to CD40LG in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0573 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of CD40 Ligand/TNFSF5
CD40 ligand (CD40L) is a type II membrane protein of 261 amino acids on activated T cells that induces B cell proliferation and immunoglobulin secretion. It has homology with tumour necrosis factor-alpha and -beta, and has important functions in B-cell activation and differentiation. Human CD40L with 5 exons, is mapped to the proximal region of the mouse X chromosome on Xq26.3-27.1, and can be detected on T cells but is absent from B cells and monocytes. Since CD40L is expressed on platelets and released from them on activation, its predictive value as a marker for clinical outcome and the therapeutic effect of inhibition of glycoprotein IIb/IIIa receptor in patients with acute coronary syndromes was investigated. The soluble CD40L may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells.CD40L plays a central role in the pathophysiology of acute coronary syndromes, and has a role in the pathogenesis of coronary artery lesions.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0573-CAP | Anti-Human CD40LG Pre-coated 96-well strip microplate | 1 |
| EK0573-ST | Human CD40LG Standard | 2 vials, 10 ng/tube |
| EK0573-DA | Human CD40LG 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.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 62.5 | 125 | 250 | 500 | 1,000 | 2,000 | 4,000 |
| O.D. | 0.025 | 0.058 | 0.104 | 0.163 | 0.341 | 0.572 | 1.122 | 1.850 |
Data Example Images
Click image to see more details
Human sCD40L PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect CD40 Ligand/TNFSF5, For serum, dilution ratio is 1:5 and concentration is 0-13741 pg/ml.
For plasma, dilution ratio is 1:1 and concentration is 0-166 pg/ml..
Some articles we found to cite concentrations of CD40 Ligand/TNFSF5 in samples: 25648034, 24964251 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human CD40 Ligand/TNFSF5/CD40LG 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) | 203 | 762 | 1128 | 198 | 762 | 1161 |
| Standard deviation | 15.42 | 58.1%8 | 56.4 | 17.02 | 61.72 | 77.78 |
| CV (%) | 7.6% | 7.4% | 5% | 8.6% | 8.1% | 6.7% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of CD40 Ligand/TNFSF5 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 | 203 | 178 | 187 | 209 | 194 | 12.35 | 6.3% |
| Sample 2 | 762 | 793 | 661 | 730 | 736 | 48.95 | 6.6% |
| Sample 3 | 1128 | 1109 | 1044 | 1131 | 1103 | 35.09 | 3.1% |
Protein Target Info & Infographic
Gene/Protein Information For CD40LG (Source: Uniprot.Org, NCBI)
Gene Name
CD40LG
Full Name
CD40 ligand
Weight
29274 MW
Superfamily
tumor necrosis factor family
Alternative Names
CD40 ligand;CD40-L;T-cell antigen Gp39;TNF-related activation protein;TRAP;Tumor necrosis factor ligand superfamily member 5;CD154;CD40 ligand, membrane form;CD40 ligand, soluble form;CD40LG;CD40L, TNFSF5, TRAP; CD40LG CD154, CD40L, HIGM1, IGM, IMD3, T-BAM, TNFSF5, TRAP, gp39, hCD40L CD40 ligand CD40 ligand|CD40 ligand|CD40-L|T-B cell-activating molecule|T-cell Gp39|TNF-related activation protein|tumor necrosis factor (ligand) superfamily member 5
*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 CD40LG, check out the CD40LG 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 CD40LG: 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 CD40 Ligand/TNFSF5/CD40LG ELISA Kit PicoKine® (EK0573)
Hello CJ!
EK0573 has been cited in 2 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.
CD40 in Retinal Müller Cells Induces P2X7-Dependent Cytokine Expression in Macrophages/Microglia in Diabetic Mice and Development of Early Experimental Diabetic Retinopathy
CD40 in Retinal M?ller Cells Induces P2X7-Dependent Cytokine Expression in Macrophages/Microglia in Diabetic Mice and Development of Early Experimental Diabetic Retinopathy
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human CD40 Ligand/TNFSF5/CD40LG ELISA Kit PicoKine®
Question
Q: Can CD40 Ligand ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
W. Zhao
Verified customer
Asked: 2020-05-01
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 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 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 view 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: 2020-05-01
Question
Q: how to proceed with the analysis of ELISA data? I have performed CD40 Ligand level in cell lysates.
Verified Customer
Verified customer
Asked: 2019-08-26
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: 2019-08-26
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-08-19
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: 2019-08-19
Question
Q: What is the optimal O.D. value for CD40 Ligand ELISA kit? I performed your CD40 Ligand 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 CD40 Ligand even though the O.D. values are not very high?
K. Baker
Verified customer
Asked: 2019-08-15
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 the above 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. 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 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 CD40 Ligand ELISA kit to have sensitivity of 15pg/ml, that means the minimum amount of CD40 Ligand that can be declared/interpreted as positive by the above standard is 15pg/ml.
Boster Scientific Support
Answered: 2019-08-15
Question
Q: if the enzyme conjugated CD40 Ligand 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 CD40 Ligand antigen?
Verified Customer
Verified customer
Asked: 2018-05-14
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: 2018-05-14
Question
Q: we need your recommendation regarding the dilution ratio of serum samples for detection of CD40 Ligand in Human cell lysates? I am trying to measure a few parameters 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: 2017-11-28
Answer
A: without knowing 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 Ligand 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: 2017-11-28
Question
Q: can you tell me how to prepare cell lysates prepared for use in Picokine® ELISA kits?
W. Surve
Verified customer
Asked: 2015-03-06
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present 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: 2015-03-06
