Human IL-12(p70) ELISA Kit PicoKine™

Human IL-12 (p70) ELISA Kit PicoKine™ (96 Tests). Quantitate Human IL12A in cell culture supernatants and serum. Sensitivity: 2pg/ml. Cited in 15 publication(s).

Product Info Summary

SKU: EK0421
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants and serum.

Product Name

Human IL-12(p70) ELISA Kit PicoKine™

See all IL12A products

SKU/Catalog Number

EK0421

Size

96 wells/kit, with removable strips.

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

Description

Human IL-12 (p70) ELISA Kit PicoKine™ (96 Tests). Quantitate Human IL12A in cell culture supernatants and serum. Sensitivity: 2pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)

Cite This Product

Human IL-12(p70) ELISA Kit PicoKine™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0421)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: (I23-S328) + (R23-S219)

Sensitivity

<2 pg/ml

Assay Range

7.8 pg/ml - 500 pg/ml

Cross Reactivity

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

Reactive Species

EK0421 is reactive to IL12A in Human samples

Validated Sample Types

cell culture supernatants and serum.

Application Guarantee

EK0421 is guaranteed for ELISA in Human by Boster Guarantee

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

Kit Components

Catalog Number Description Quantity
EK0421-CAP Anti-Human IL12A Pre-coated 96-well strip microplate 1
EK0421-ST Human IL12A Standard 2 vials, 10 ng/tube
EK0421-DA Human IL12A 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.
  • 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)07.815.631.362.5125250500
O.D.0.0190.0690.1260.2390.4410.8341.4412.086

Data Example Images

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human 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 PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)18512941955310
Standard deviation1.313.4117.381.673.7921.39
CV (%)7.3%6.7%5.2%8.8%6.9%6.9%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of IL12A 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 118182017181.086%
Sample 251596054563.676.5%
Sample 32942902842962914.581.5%
*number of samples for each test n=16.

Gene/Protein Information For IL12A (Source: Uniprot.Org, NCBI)

Uniprot ID

P29459

Gene ID

3592

Gene Name

IL12A

Full Name

Interleukin-12 subunit alpha

Weight

24.874kDa

Superfamily

IL-6 superfamily

Alternative Names

CLMF p35; Cytotoxic lymphocyte maturation factor 35 kDa subunit; IL12 p70; IL-12 p70; IL-12 subunit p35; IL-12, subunit p35; interleukin 12, p35; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35); interleukin-12 alpha chain; interleukin-12 subunit alpha; natural killer cell stimulatory factor 1, 35 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NKSF1; p35 IL12A CLMF, IL-12A, NFSK, NKSF1, P35 interleukin 12A interleukin-12 subunit alpha|CLMF p35|IL-12, subunit p35|IL35 subunit|NF cell stimulatory factor chain 1|NK cell stimulatory factor chain 1|cytotoxic lymphocyte maturation factor 1, p35|cytotoxic lymphocyte maturation factor 35 kDa subunit|interleukin 12, p35|interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocyte maturation factor 1, p35)|interleukin-12 alpha chain

*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 IL12A, check out the IL12A Infographic

IL12A 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 IL12A: 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]

EK0421 has been cited in 15 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.

Singh A,Das D,Kurra S,Arava S,Gupta S,Sharma A.Dendritic cells and their associated pro-inflammatory cytokines augment to the inflammatory milieu in vitiligo skin.Cytokine.2021 Jun 5:155598.doi:10.1016/j.cyto.2021.155598.Epub ahead of print.PMID:34103210.
Species: Human
EK0421 usage in article: APP:ELISA, SAMPLE:DENDRITIC CELL, DILUTION:NA

Das D, Singh A, Antil PS, Sharma D, Arava S, Khandpur S, Sharma A. Distorted frequency of dendritic cells and their associated stimulatory and inhibitory markers augment the pathogenesis of pemphigus vulgaris. Immunol Res. 2020 Nov 12. doi: 10.1007/s12026
Species: Human
EK0421 usage in article: APP:ELISA, SAMPLE:PBMCS, DILUTION:NA

Elevated serum neopterin levels in children with functional constipation: association with systemic proinflammatory cytokines

Interleukin-12 and interferon-γ acting on damp-heat of spleen- stomach syndrome triggered by Helicobacter pylori

The influence of interferon-β supplemented human dendritic cells on BCG immunogenicity

Pancreatic cancer derived exosomes regulate the expression of TLR4 in dendritic cells via miR-203

Gui J, Xiong F, Yang W, Li J, Huang G. Am J Reprod Immunol. 2012 May;67(5):383-90. Doi: 10.1111/J.1600-0897.2011.01097.X. Epub 2012 Jan 9. Effects Of Acupuncture On Lif And Il-12 In Rats Of Implantation Failure.

Li Y, Li X, Liu H, Zhuang S, Yang J, Zhang F. Oncol Lett. 2014 Feb;7(2):576-582. Epub 2013 Dec 9. Intranasal Immunization With Recombinant Lactococci Carrying Human Papillomavirus E7 Protein And Mouse Interleukin-12 Dna Induces E7-Specific Antitum...

Hara T, Mimura K, Abe T, Shioi G, Seiki M, Sakamoto T. J Biol Chem. 2011 Sep 16;286(37):32542-51. Doi: 10.1074/Jbc.M111.271726. Epub 2011 Jul 21. Deletion Of The Mint3/Apba3 Gene In Mice Abrogates Macrophage Functions And Increases Resistance To L...

Li P, Yu Q, Ye X, Wang Z, Yang Q. Microbiology. 2011 Sep;157(Pt 9):2639-46. Doi: 10.1099/Mic.0.049148-0. Epub 2011 Jun 23. Lactobacillus S-Layer Protein Inhibition Of Salmonella-Induced Reorganization Of The Cytoskeleton And Activation Of Mapk Sig...

Ma Yj, He M, Han Ja, Yang L, Ji Xy. Scand J Immunol. 2013 Oct;78(4):387-93. Doi: 10.1111/Sji.12097. A Clinical Study Of Hbsag-Activated Dendritic Cells And Cytokine-Induced Killer Cells During The Treatment For Chronic Hepatitis B.

Li Y, Li Sy, Li Js, Deng L, Tian Yg, Jiang Sl, Wang Y, Wang Yy. Biol Pharm Bull. 2012;35(10):1752-60. Epub 2012 Aug 3. A Rat Model For Stable Chronic Obstructive Pulmonary Disease Induced By Cigarette Smoke Inhalation And Repetitive Bacterial Infe...

Li J, Li Ff, Zuo W, Zhou Y, Hao Hy, Dang J, Jiang M, He Mz, Deng Dr. J Huazhong Univ Sci Technolog Med Sci. 2014 Aug;34(4):586-90. Doi: 10.1007/S11596-014-1320-2. Epub 2014 Aug 19. Up-Regulated Expression Of Tim-3/Gal-9 At Maternal-Fetal Interface...

Yu Zm, Liu Zh, Chen J, Zeng Q. Chin J Integr Med. 2014 Dec;20(12):934-43. Doi: 10.1007/S11655-014-1863-X. Epub 2014 Nov 27. Anti-Inflammatory Effect Of Qingwen Baidu Decoction (????????????

Pan Zy, Wang H. Int Immunopharmacol. 2014 May;20(1):229-37. Doi: 10.1016/J.Intimp.2014.03.004. Epub 2014 Mar 19. Synergistic Interaction Between Choline And Aspirin Against Acute Inflammation Induced By Carrageenan And Lipopolysaccharide.

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13 Customer Q&As for Human IL-12(p70) ELISA Kit PicoKine™

Question

Q: What is the optimal O.D. value for IL12A ELISA kit? I used 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?

W. Carter

Verified customer

Asked: 2020-12-11

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. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. in the example mentioned, 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 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 IL12A ELISA kit to have sensitivity of 2pg/ml, that means the minimum amount of IL12A that can be declared/interpreted as positive by the above standard is 2pg/ml.

Boster Scientific Support

Answered: 2020-12-11

Question

Q: how to thaw whole blood sample for IL12A ELISA after freezing?

G. Smith

Verified customer

Asked: 2020-11-10

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 separate the serum to freeze for later analysis.

Boster Scientific Support

Answered: 2020-11-10

Question

I recently purchased the EK0421 Human IL-12(p70) ELISA Kit PicoKine. The results of the standard curve are fine however, all of the cytokines are below the curve. Any suggestions about it? I was wondering if concentrating the cytokines in the samples or adding more volume would help. What do you think?

Verified Customer

Verified customer

Asked: 2019-12-03

Answer

In cases of low signal, it might be the result of incubation time. We would suggest that you increased your incubation period of assay to overnight. Also we recommend to prepared all reagents as you use them, especially the HRP-Streptavidin solution. The concentration of the cytokines or an increase in the volumes of sample used maybe also effective if these other steps fail to increase signal. But, the minimum detectable dose of EK0421 Human IL-12(p70) is typically less than 1 pg/ml.

Boster Scientific Support

Answered: 2019-12-03

Question

Regarding your ELISA kit EK0421, can you please advise whether this kit was designed to specifically target the p70 heterodimer, or if it detects any of the other single subunits?

Verified Customer

Verified customer

Asked: 2019-11-29

Answer

EK0421 detects the p70 heterodimer only.

Boster Scientific Support

Answered: 2019-11-29

Question

Q: Can IL12A ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2019-10-30

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 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 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 check 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: 2019-10-30

Question

Does EK0421 kit detect other single subunits as well?

Verified customer

Asked: 2019-07-22

Answer

The Human IL-12(p70) ELISA Kit PicoKine EK0421 detects the p70 heterodimer only.

Boster Scientific Support

Answered: 2019-07-22

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

A. Thompson

Verified customer

Asked: 2019-06-30

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: 2019-06-30

Question

Does EK0421 detect the p70 heterodimer or does it also detect other single subunits?

Verified Customer

Verified customer

Asked: 2019-06-06

Answer

EK0421 detects the p70 heterodimer only.

Boster Scientific Support

Answered: 2019-06-06

Question

Q: how to analyze ELISA data? I measured IL12A level in serum.

Verified Customer

Verified customer

Asked: 2018-09-28

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: 2018-09-28

Question

Q: if the enzyme conjugated IL12A 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 IL12A antigen?

D. Wilson

Verified customer

Asked: 2018-08-07

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: 2018-08-07

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis IL12A using the EK0421 Human IL12A Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2018-03-10

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to prevent protein degradation.

Boster Scientific Support

Answered: 2018-03-10

Question

Q: can you recommend the dilution ratio of serum samples for detection of IL12A in Human serum? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2017-12-03

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: 2017-12-03

Question

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

Verified Customer

Verified customer

Asked: 2016-11-22

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: 2016-11-22

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