Human CD10/Neprilysin ELISA Kit PicoKine®

Neprilysin/CD10 ELISA kit for Human

Human CD10/Neprilysin ELISA Kit PicoKine™ (96 Tests). Quantitate Human MME in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK0851
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Human CD10/Neprilysin ELISA Kit PicoKine®

View all Neprilysin/CD10 ELISA kits

SKU/Catalog Number

EK0851

Size

96 wells/kit, with removable strips.

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

Description

Human CD10/Neprilysin ELISA Kit PicoKine™ (96 Tests). Quantitate Human MME in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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 CD10/Neprilysin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0851)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: CHO; Immunogen sequence: Y52-W750

Sensitivity

<10 pg/ml

Assay Range

125 pg/ml - 8,000 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 EK0851 for more details

Cross-reactivity

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

Reactive Species

EK0851 is reactive to MME in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK0851 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of Neprilysin/CD10

CD10, also known as membrane metallo-endopeptidase, neutral endopeptidase (NEP), Neprilysin, or common acute lymphoblastic leukemia antigen (CALLA), is a zinc-dependent metalloprotease enzyme that degrades a number of small secreted peptides, most notably theamyloid beta peptide whose abnormal misfolding and aggregation in neural tissue has been implicated as a cause of Alzheimer's disease. This gene is localized to human chromosome 3 by study of somatic cell hybrids and regionalized the location to 3q21-q27 by in situ hybridization. By cDNA transfection analysis, CD10 is confirmed as a functional neutral endopeptidase of the type that has previously been called enkephalinase. CD10 has also been called atriopeptidase. Atriopeptidase specifically degrades atrial natriuretic factor. A specific enzyme inhibitor was developed and reported that it had effects similar to those of low-dose ANF infusion. These effects include diuresis, natriuresis, vasodilatation, and suppression of the renin-angiotensin-aldosterone system.

Kit Components

Catalog Number Description Quantity
EK0851-CAP Anti-Human MME Pre-coated 96-well strip microplate 1
EK0851-ST Human MME Standard 2 vials, 10 ng/tube
EK0851-DA Human MME 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)01252505001000200040008000
O.D.0.0970.1480.1960.3190.4620.8581.3281.965

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Neprilysin/CD10, Dilution ratio of 1:1, concentration in serum and plasma is around 1 ng/ml..

Some articles we found to cite concentrations of Neprilysin/CD10 in samples: 24972738 (Pubmed IDs).

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human CD10/Neprilysin 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)201216643162168814302
Standard deviation14.8751.62233.0617.9260.78305.44
CV (%)7.4%5.4%5.4%8.3%6.9%7.1%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Neprilysin/CD10 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 120117018118118311.186.1%
Sample 2956105810111064102243.44.2%
Sample 34316449045324435444381.121.8%
*number of samples for each test n=16.

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

Gene Name

MME

Full Name

Neprilysin

Weight

85.514kDa

Superfamily

peptidase M13 family

Alternative Names

Atriopeptidase; CALLA; CALLAmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase); CD10 antigen; CD10; CD10); CD10membrane metallo-endopeptidase variant 1; Common acute lymphocytic leukemia antigen; DKFZp686O16152; EC 3.4.24; EC 3.4.24.11; Enkephalinase; EPN; Leu-19; membrane metallo-endopeptidase variant 2; membrane metallo-endopeptidase; MGC126681; MGC126707; MME; NEPmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA; Neprilysin; Neutral Endopeptidase 24.11; Neutral endopeptidase; NKH1; SFE; Skin fibroblast elastase MME CALLA, CD10, CMT2T, NEP, SCA43, SFE membrane metalloendopeptidase neprilysin|atriopeptidase|common acute lymphocytic leukemia antigen|membrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA, CD10)|membrane metallo-endopeptidase variant 1|membrane metallo-endopeptidase variant 2|neprilysin-390|neprilysin-411|neutral endopeptidase 24.11|skin fibroblast elastase

*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 MME, check out the MME Infographic

MME 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 MME: 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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9 Customer Q&As for Human CD10/Neprilysin ELISA Kit PicoKine®

Question

Q: how can I thaw whole blood sample for CD10 ELISA after freezing?

Verified Customer

Verified customer

Asked: 2020-10-22

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 CD10 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: 2020-10-22

Question

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

P. Allen

Verified customer

Asked: 2020-07-17

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: 2020-07-17

Question

Q: how do I analyze ELISA data? I have obtained CD10 level in serum.

V. Sunder

Verified customer

Asked: 2020-03-01

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: 2020-03-01

Question

Q: Can CD10 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

A. Patel

Verified customer

Asked: 2020-01-30

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 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 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: 2020-01-30

Question

Q: What is the optimal O.D. value for CD10 ELISA kit? I used your CD10 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. is it safe to say these samples contain CD10 even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2019-10-24

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. considering your assay, 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. typically, 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 CD10 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of CD10 that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2019-10-24

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring CD10 using the EK0851 Human CD10 Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-10-01

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 on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples in order to limit protein degradation.

Boster Scientific Support

Answered: 2019-10-01

Question

Q: if the enzyme conjugated CD10 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 CD10 antigen?

Verified Customer

Verified customer

Asked: 2019-06-12

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: 2019-06-12

Question

Q: can I use citrate plasma as samples in Human CD10 Picokine® ELISA Kit (Catalog # EK0851)?

Verified Customer

Verified customer

Asked: 2018-06-29

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of CD10 causing disruption of its protein structure. CD10 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 CD10 ELISA, treating samples with a number of anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2018-06-29

Question

Q: can you recommend the dilution ratio of serum samples for detection of CD10 in Human serum? I am trying to measure a a number of 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: 2018-04-20

Answer

A: unable to know 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 CD10 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: 2018-04-20

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