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Product Info Summary
| SKU: | EK0423 |
|---|---|
| Size: | 96wells/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 IL-12(p40) ELISA Kit PicoKine®
SKU/Catalog Number
EK0423
Size
96wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human IL-12 (p40) ELISA Kit PicoKine® (96 Tests). Quantitate Human IL12B in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 2pg/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 IL-12(p40) ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0423)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: sf21; Immunogen sequence: I23-S328
Capture/Detection Antibodies
monoclonal antibody from mouse|polyclonal antibody from goat
Sensitivity
<2pg/ml
Assay Range
31.2pg/ml-2000pg/ml
Standard Dilution Instructions
See datasheet of EK0423 for more details
Cross-reactivity
There is cross-reactivity with IL-12(p70), IL-23(p19/p40)< 5%.
Reactive Species
EK0423 is reactive to IL12B in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
Application Guarantee
EK0423 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of IL12B
Interleukin-12 (IL12; formerly NKSF, for natural killer cell stimulatory factor, or CLMF, for cytotoxic lymphocyte maturation factor) is a novel cytokine cloned from B-cell lines. IL-12 is a heterodimeric molecule composed of p35 and p40 subunits. The larger 40-kDa subunit (p40) is a member of the cytokine receptor family, and the smaller 35-kDa subunit (p35) is related to IL6 and GCSF. Both IL-12 p40 (-/-) and p35 (-/-) mice fail to produce IL-12 p70 heterodimer. Interleukin (IL)-12 has been cloned on the basis of its ability to activate natural killer (NK) cells and promote the development of cytolytic T cells. With further understanding of its activities, IL-12 has emerged as an important cytokine, affecting both immune and hematologic functions. It has been shown to be necessary for the T cell independent induction of interferon (IFN)-gamma, critical for the initial suppression of bacterial and parasitic infection; for the development of a Th1 response, critical for effective host defense against intracellular pathogens; and for the activation of differentiated T lymphocytes of both CD4+ and CD8+ phenotype. The subunits map to different chromosomes: p40 (IL12B) to 5q31-q33 and p35 (IL12A) to 3p12-3q13.2.2 The standard product used in this kit is recombinant human IL-12 (p40), which is a 40KDa glycoprotein consisting of 306 amino acids.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0423-CAP | Anti-Human IL12B Pre-coated 96-well strip microplate | 1 |
| EK0423-ST | Human IL12B Standard | 2 vials, 2ng/tube |
| EK0423-DA | Human IL12B 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
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.029 | 0.135 | 0.218 | 0.400 | 0.753 | 1.253 | 1.884 | 2.567 |
Data Example Images
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Human IL-12(p40) PicoKine ELISA Kit standard curve
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human IL-12(p40) 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) | 75 | 257 | 881 | 75 | 271 | 808 |
| Standard deviation | 4.5 | 13.1 | 63.43 | 6%5 | 18.69 | 71.1 |
| CV (%) | 6% | 8.8%% | 7.2% | 7% | 6.9% | 8.8% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of IL12B in ELISA kits from four different production batches/lots.
| Lots | Lot1 (pg/ml) | Lot2 (pg/ml) | Lot3 (pg/ml) | Lot4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 75 | 75 | 66 | 64 | 70 | 5.04 | 7.2% |
| Sample 2 | 257 | 230 | 241 | 245 | 243 | 9.65 | 3.9% |
| Sample 3 | 881 | 962 | 909 | 827 | 894 | 48.74 | 5.4% |
Specific Publications For Human IL-12(p40) ELISA Kit PicoKine® (EK0423)
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human IL-12(p40) ELISA Kit PicoKine®
Question
I purchased EK0423 antibody, however I have some queries; is the kit be use with tissue homogenate? My tissue prepared by mechanical lysis and homogenate buffer contains 0.5%Tween 20, EDTA and Tris. Would the detergent interfere with the assay?
Verified Customer
Verified customer
Asked: 2019-12-20
Answer
The product EK0423 antibody is suitable for tissue homogenates and the suggested homogenate buffer containing 0.5% Tween20, EDTA and Tris is appropriate with these kit.
Boster Scientific Support
Answered: 2019-12-20
Question
Q: Can Il12b ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2019-09-12
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 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: 2019-09-12
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2019-08-11
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 needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2019-08-11
Question
Q: can you recommend the dilution ratio of serum samples for detection of Il12b in Human serum? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.
B. Moore
Verified customer
Asked: 2019-04-02
Answer
A: unable to understand 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 Il12b 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: 2019-04-02
Question
Q: how to proceed with the analysis of ELISA data? I measured Il12b level in serum.
Verified Customer
Verified customer
Asked: 2018-12-10
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: 2018-12-10
Question
Q: how can I thaw whole blood sample for Il12b ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-06-07
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 Il12b 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: 2018-06-07
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
K. Yang
Verified customer
Asked: 2017-04-28
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 affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2017-04-28
