Human ADA/Adenosine Deaminase ELISA Kit PicoKine®

Adenosine Deaminase/ADA ELISA kit for Human

Human ADA/Adenosine Deaminase ELISA Kit PicoKine™ (96 Tests). Quantitate Human ADA in cell culture supernatants, serum and pleural fluid. Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1446
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and pleural fluid.

Product Name

Human ADA/Adenosine Deaminase ELISA Kit PicoKine®

View all Adenosine Deaminase/ADA ELISA kits

SKU/Catalog Number

EK1446

Size

96 wells/kit, with removable strips.

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

Description

Human ADA/Adenosine Deaminase ELISA Kit PicoKine™ (96 Tests). Quantitate Human ADA in cell culture supernatants, serum and pleural fluid. Sensitivity: 10pg/ml.

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 ADA/Adenosine Deaminase ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1446)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: A2-L363

Sensitivity

<10 pg/ml

Assay Range

31.2 pg/ml - 2,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1446 for more details

Cross-reactivity

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

Reactive Species

EK1446 is reactive to ADA in Human samples

Validated Sample Types

cell culture supernatants, serum and pleural fluid.

Application Guarantee

EK1446 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of Adenosine Deaminase/ADA

Adenosine Deaminase (also known as adenosine aminohydrolase, or ADA) is an enzyme involved in purine metabolism. It is needed for the breakdown of adenosine from food and for the turnover of nucleic acids in tissues. ADA in humans is involved in the development and maintenance of the immune system. However, ADA association has also been observed with epithelial cell differentiation, neurotransmission, and gestation maintenance. It has also been proposed that ADA, in addition to adenosine breakdown, stimulates release of excitatory amino acids and is necessary to the coupling of A1 adenosine receptors and heterotrimeric G proteins. Adenosine deaminase deficiency leads to pulmonary fibrosis, suggesting that chronic exposure to high levels of adenosine can exacerbate inflammation responses rather than suppressing them. It has also been recognized that adenosine deaminase protein and activity is upregulated in mouse hearts that overexpress HIF-1 alpha, which in part explains the attenuated levels of adenosine in HIF-1 alpha expressing hearts during ischemic stress.

Kit Components

Catalog Number Description Quantity
EK1446-CAP Anti-Human ADA Pre-coated 96-well strip microplate 1
EK1446-ST Human ADA Standard 2 vials, 10 ng/tube
EK1446-DA Human ADA 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.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)031.262.512525050010002000
O.D.0.0210.0780.1240.2270.3980.7121.2421.802

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Adenosine Deaminase/ADA, Dilution ratio of 1:1, concentration in serum is around 400 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human ADA/Adenosine Deaminase 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)762571026812401095
Standard deviation4.6312.3341.044.0111.5253.65
CV (%)6.1%4.8%4%7.3%4.8%4.9%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Adenosine Deaminase/ADA 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 176696470694.266.1%
Sample 22572302402462439.784%
Sample 3102687889595493857.986.1%
*number of samples for each test n=16.

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

Gene Name

ADA

Full Name

Adenosine deaminase

Weight

40.764kDa

Superfamily

metallo-dependent hydrolases superfamily

Alternative Names

ADA; ADA1; adenine deaminase; Adenosine aminohydrolase; Adenosine Deaminase; EC 3.5.4.4 ADA ADA1 adenosine deaminase adenosine deaminase|adenosine aminohydrolase

*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 ADA, check out the ADA Infographic

ADA 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 ADA: 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].

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7 Customer Q&As for Human ADA/Adenosine Deaminase ELISA Kit PicoKine®

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring Adenosine Deaminase using the EK1446 Human Adenosine Deaminase Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2020-01-11

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may affect 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 repetitive freeze-thaw cycles with the stored samples to limit protein degradation.

Boster Scientific Support

Answered: 2020-01-11

Question

What is the reference range for normal ranges of ADA concentration when using EK1446?

Verified Customer

Verified customer

Asked: 2019-03-29

Answer

Normal ranges of ADA concentration when using EK1446 is 200-500pg/mL

Boster Scientific Support

Answered: 2019-03-29

Question

Q: is it okay to use heparin plasma as samples in Human Adenosine Deaminase Picokine® ELISA Kit (Catalog # EK1446)?

S. Dayal

Verified customer

Asked: 2018-12-29

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of Adenosine Deaminase causing disruption of its protein structure. Adenosine Deaminase 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 Adenosine Deaminase ELISA, treating samples with various 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: 2018-12-29

Question

Q: which procedure should I follow in order to thaw whole blood sample for Adenosine Deaminase ELISA after freezing?

Verified Customer

Verified customer

Asked: 2018-04-07

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 Adenosine Deaminase 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: 2018-04-07

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

W. Hill

Verified customer

Asked: 2017-11-02

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: 2017-11-02

Question

Q: how do I analyze ELISA data? I have performed Adenosine Deaminase level in pleural fluid.

Verified Customer

Verified customer

Asked: 2016-10-26

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: 2016-10-26

Question

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

Verified Customer

Verified customer

Asked: 2015-07-05

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 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 see 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: 2015-07-05

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