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Product Info Summary
| SKU: | EK1652 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Rat |
| Application: | ELISA |
| Sample Types: | cell culture supernatants and serum. |
Product info
Product Name
Rat IL-12(p70) ELISA Kit PicoKine®
SKU/Catalog Number
EK1652
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Rat IL-12(p70) ELISA Kit PicoKine® (96 Tests). Quantitate Rat IL-12(p70) in cell culture supernatants and serum. Sensitivity: 1.5pg/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
Rat IL-12(p70) ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1652)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: (M23 - S335) + (R23 - S215)
Sensitivity
<1.5 pg/ml
Assay Range
15.6 pg/ml - 1,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1652 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1652 is reactive to IL-12(p70) in Rat samples
Validated Sample Types
cell culture supernatants and serum.
Application Guarantee
EK1652 is guaranteed for ELISA in Rat by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Il12a
Interleukin (IL)-12 is a 70-KDa cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response. And the IL-12p70 is a heterodimer of p35 and p40 subunits; it is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation. Gene expression analysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40 (protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT- 18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70. The standard product used in this kit is recombinant rat IL-12 p70 comprising of a 193 amino acids p35 chain and a 313 amino acids p40 chain connected by a disulfide bond.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1652-CAP | Anti-Rat IL-12(p70) Pre-coated 96-well strip microplate | 1 |
| EK1652-ST | Rat IL-12(p70) Standard | 2 vials, 10 ng/tube |
| EK1652-DA | Rat IL-12(p70) 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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.019 | 0.065 | 0.121 | 0.214 | 0.384 | 0.632 | 1.020 | 1.813 |
Data Example Images
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Rat IL-12(p70) PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Il12a, Dilution ratio of 1:10, concentration in serum is 600-3200 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Rat IL-12(p70) 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) | 35 | 222 | 502 | 34 | 224 | 454 |
| Standard deviation | 1.92 | 15.76 | 27.61 | 2.07 | 19.93 | 29.05 |
| CV (%) | 5.5% | 7.1% | 5.5% | 6.1% | 8.9% | 6.4% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Il12a 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 | 35 | 31 | 36 | 34 | 34 | 1.87 | 5.5% |
| Sample 2 | 222 | 226 | 219 | 229 | 224 | 3.8 | 1.6% |
| Sample 3 | 502 | 447 | 478 | 479 | 476 | 19.55 | 4.1% |
Protein Target Info & Infographic
Gene/Protein Information For IL-12(p70) (Source: Uniprot.Org, NCBI)
Gene Name
IL-12(p70)
Full Name
Interleukin-12 subunit alpha
Weight
24.246kDa
Superfamily
IL-6 superfamily
Alternative Names
Interleukin-12 subunit alpha; IL-12A; Cytotoxic lymphocyte maturation factor 35 kDa subunit; CLMF p35; IL-12 subunit p35; Il12a Il12a|interleukin 12A|interleukin-12 subunit alpha|CLMF p35|IL-12 subunit p35|IL-12A|cytotoxic lymphocyte maturation factor 35 kDa subunit|interleukin 12 p35 subunit
*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 IL-12(p70), check out the IL-12(p70) 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 IL-12(p70): 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 Rat IL-12(p70) ELISA Kit PicoKine® (EK1652)
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Customer Reviews
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Customer Q&As
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Find answers in Q&As, reviews.
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12 Customer Q&As for Rat IL-12(p70) ELISA Kit PicoKine®
Question
Q: how to thaw whole blood sample for IL12A ELISA after freezing?
P. Gandhi
Verified customer
Asked: 2020-12-13
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 IL12A 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: 2020-12-13
Question
Q: how to analyze ELISA data? I have obtained IL12A level in serum.
J. Taylor
Verified customer
Asked: 2020-11-11
Answer
A: please read 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: 2020-11-11
Question
Q: is it okay to use EDTA plasma as samples in Rat IL12A Picokine® ELISA Kit (Catalog # EK1652)?
Verified Customer
Verified customer
Asked: 2020-04-06
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of IL12A causing degradation of its protein structure. IL12A may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the IL12A ELISA, treating samples with a number of anticoagulants and decided that / can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2020-04-06
Question
Q: Can IL12A ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
V. Brown
Verified customer
Asked: 2020-03-12
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: 2020-03-12
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
K. Perez
Verified customer
Asked: 2019-08-31
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: 2019-08-31
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2019-06-03
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are included in the product insert. Components in lysate and lysis buffer can affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2019-06-03
Question
Q: What is the optimal O.D. value for IL12A ELISA kit? I performed your IL12A 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 IL12A even though the O.D. values are not very high?
L. Kapoor
Verified customer
Asked: 2019-04-03
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. 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 relation 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 IL12A ELISA kit to have sensitivity of 1.5pg/ml, that means the minimum amount of IL12A that can be declared/interpreted as positive by the above standard is 1.5pg/ml.
Boster Scientific Support
Answered: 2019-04-03
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2018-12-25
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 vary slightly by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2018-12-25
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of IL12A in Rat serum? I am trying to measure a multiple 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.
Verified Customer
Verified customer
Asked: 2018-06-18
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 IL12A 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: 2018-06-18
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
C. King
Verified customer
Asked: 2017-11-13
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: 2017-11-13
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring IL12A using the EK1652 Rat IL12A Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2016-07-16
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample right after collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples to prevent protein degradation.
Boster Scientific Support
Answered: 2016-07-16
Question
Q: if the enzyme conjugated IL12A 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 IL12A antigen?
Verified Customer
Verified customer
Asked: 2015-03-04
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: 2015-03-04
