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Product Info Summary
| SKU: | EK2052 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA, citrate). |
Product info
Product Name
Human CD69 ELISA Kit PicoKine®
SKU/Catalog Number
EK2052
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human CD69 ELISA Kit PicoKine® (96 Tests). Quantitate Human CD69 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 50pg/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 CD69 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK2052)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: S62-K199
Sensitivity
<10 pg/ml
Assay Range
31.2 pg/ml - 2,000 pg/ml
Standard Dilution Instructions
See datasheet of EK2052 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK2052 is reactive to CD69 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
Application Guarantee
EK2052 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of CD69
CD69 (Cluster of Differentiation 69) is a human transmembrane C-Type lectin protein encoded by the CD69 gene. This gene is mapped to 6 F3; 6 63.21 cM. This gene encodes a member of the calcium dependent lectin superfamily of type II transmembrane receptors. Expression of the encoded protein is induced upon activation of T lymphocytes, and may play a role in proliferation. Furthermore, the protein may act to transmit signals in natural killer cells and platelets.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK2052-CAP | Anti-Human CD69 Pre-coated 96-well strip microplate | 1 |
| EK2052-ST | Human CD69 Standard | 2 vials, 10 ng/tube |
| EK2052-DA | Human CD69 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 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.031 | 0.129 | 0.217 | 0.389 | 0.759 | 1.223 | 2.057 | 2.432 |
Data Example Images
Click image to see more details
Human CD69 PicoKine ELISA Kit Standard Curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect CD69, Dilution ratio of 1:1, concentration in serum and plasma is 20-144 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human CD69 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) | 109 | 470 | 1804 | 112 | 481 | 1927 |
| Standard deviation | 5.23 | 23.03 | 82.98 | 6.27 | 33.19 | 123.33 |
| CV (%) | 4.8% | 4.9% | 4.6% | 5.6% | 6.9% | 6.4% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of CD69 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 | 109 | 106 | 115 | 107 | 109 | 3.49 | 3.2% |
| Sample 2 | 470 | 422 | 410 | 478 | 445 | 29.44 | 6.6% |
| Sample 3 | 1804 | 1954 | 1992 | 1834 | 1896 | 83.42 | 4.4% |
Protein Target Info & Infographic
Gene/Protein Information For CD69 (Source: Uniprot.Org, NCBI)
Gene Name
CD69
Full Name
Early activation antigen CD69
Weight
22.559kDa
Alternative Names
Early activation antigen CD69; Activation inducer molecule; AIM; BL-AC/P26; C-type lectin domain family 2 member C; EA1; Early T-cell activation antigen p60; GP32/28; Leukocyte surface antigen Leu-23; MLR-3; CD69; CLEC2C CD69 AIM, BL-AC/P26, CLEC2C, EA1, GP32/28, MLR-3 CD69 molecule early activation CD69|C-type lectin domain family 2, member C|CD69 (p60, early T-cell activation )|activation inducer molecule (AIM/CD69)|early T-cell activation p60|early lymphocyte activation |leukocyte surface Leu-23
*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 CD69, check out the CD69 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 CD69: 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 CD69 ELISA Kit PicoKine® (EK2052)
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No publications found for EK2052
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Customer Reviews
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Customer Q&As
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10 Customer Q&As for Human CD69 ELISA Kit PicoKine®
Question
Q: Can CD69 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
C. Gonzalez
Verified customer
Asked: 2020-08-18
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 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: 2020-08-18
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2020-05-19
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 a must for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2020-05-19
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2020-01-28
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 affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2020-01-28
Question
Q: which procedure should I follow in order to thaw whole blood sample for CD69 ELISA after freezing?
Verified Customer
Verified customer
Asked: 2019-10-18
Answer
A: we do not recommend freezing 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 CD69 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: 2019-10-18
Question
Q: What is the optimal O.D. value for CD69 ELISA kit? I performed your CD69 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 CD69 even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-01-22
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. what you should focus on 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 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 CD69 ELISA kit to have sensitivity of 50pg/ml, that means the minimum amount of CD69 that can be declared/interpreted as positive by the above standard is 50pg/ml.
Boster Scientific Support
Answered: 2019-01-22
Question
Q: can you suggest the dilution ratio of serum samples for detection of CD69 in Human plasma? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.
B. Zhao
Verified customer
Asked: 2018-08-18
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 CD69 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: 2018-08-18
Question
Q: if the enzyme conjugated CD69 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 CD69 antigen?
Verified Customer
Verified customer
Asked: 2018-03-05
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: 2018-03-05
Question
Q: how to proceed with the analysis of ELISA data? I have performed CD69 level in plasma.
W. Bhatt
Verified customer
Asked: 2017-07-12
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: 2017-07-12
Question
Q: is it okay to use citrate plasma as samples in Human CD69 Picokine® ELISA Kit (Catalog # EK2052)?
T. Wu
Verified customer
Asked: 2016-06-07
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of CD69 causing disruption of its protein structure. CD69 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the CD69 ELISA, treating samples with different anticoagulants and decided that heparin, EDTA or citrate can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2016-06-07
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2016-03-21
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 depends upon the kit used so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2016-03-21
