Human Myeloblastin ELISA Kit PicoKine®

Proteinase 3/Myeloblastin/PRTN3 ELISA kit for Human

Human Myeloblastin ELISA Kit PicoKine™ (96 Tests). Quantitate Human PRTN3 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10 pg/ml.

Product Info Summary

SKU: EK2034
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin, EDTA).

Product Name

Human Myeloblastin ELISA Kit PicoKine®

View all Proteinase 3/Myeloblastin/PRTN3 ELISA kits

SKU/Catalog Number

EK2034

Size

96 wells/kit, with removable strips.

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

Description

Human Myeloblastin ELISA Kit PicoKine™ (96 Tests). Quantitate Human PRTN3 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10 pg/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 Myeloblastin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK2034)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: I28-R248

Sensitivity

<10 pg/ml

Assay Range

62.5 pg/ml - 4,000 pg/ml

Cross-reactivity

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

Reactive Species

EK2034 is reactive to PRTN3 in Human samples

Validated Sample Types

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

Application Guarantee

EK2034 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of Proteinase 3/Myeloblastin/PRTN3

Proteinase 3 also known as PRTN3 is an enzyme that in humans is encoded by the PRTN3 gene. This gene is mapped to 19p13.3. PRTN3 is a serine protease enzyme expressed mainly in neutrophil granulocytes. Its exact role in the function of the neutrophil is unknown, but, in human neutrophils, proteinase 3 contributes to the proteolytic generation of antimicrobial peptides. It is also the target of anti-neutrophil cytoplasmic antibodies (ANCAs) of the c-ANCA (cytoplasmic subtype) class, a type of antibody frequently found in the disease granulomatosis with polyangiitis.

Kit Components

Catalog Number Description Quantity
EK2034-CAP Anti-Human PRTN3 Pre-coated 96-well strip microplate 1
EK2034-ST Human PRTN3 Standard 2 vials, 4 ng/tube
EK2034-DA Human PRTN3 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)062.5125250500100020004000
O.D.0.0200.0890.1510.2510.4890.8621.6532.480

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Proteinase 3/Myeloblastin/PRTN3, Dilution ratio of serum 1:100, plasma 1:300, concentration in serum is around 100ng/ml, plasma around 300ng/ml.

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human Myeloblastin 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)10932615131164011765
Standard deviation4.5814.0262.037.6627.27125.32
CV (%)4.2%4.3%4.1%6.6%6.8%7.1%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Proteinase 3/Myeloblastin/PRTN3 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 11091151061131103.493.1%
Sample 232634533131732910.133.0%
Sample 31513134513721433141564.564.5%
*number of samples for each test n=16.

Gene/Protein Information For PRTN3 (Source: Uniprot.Org, NCBI)

Uniprot ID

P24158

Gene ID

5657

Gene Name

PRTN3

Full Name

Myeloblastin

Weight

27.807kDa

Superfamily

peptidase S1 family

Alternative Names

ACPA; AGP7; AGP7PR3; azurophil granule protein 7; C-ANCA; EC 3.4.21; EC 3.4.21.76; Leukocyte proteinase 3; MBN; MBT; Myeloblastin; Neutrophil proteinase 4; NP4; NP-4; P29C-ANCA antigen; PR-3CANCA; Proteinase 3; PRTN3; serine proteinase, neutrophil; Wegener autoantigen; Wegener granulomatosis autoantigen PRTN3 ACPA, AGP7, C-ANCA, CANCA, MBN, MBT, NP-4, NP4, P29, PR-3, PR3 proteinase 3 myeloblastin|C-ANCA antigen|Wegener granulomatosis autoantigen|azurophil granule protein 7|leukocyte proteinase 3|neutrophil proteinase 4|serine proteinase, neutrophil|wegener autoantigen

*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 PRTN3, check out the PRTN3 Infographic

PRTN3 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 PRTN3: 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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8 Customer Q&As for Human Myeloblastin ELISA Kit PicoKine®

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring Proteinase 3 using the EK2034 Human Proteinase 3 Picokine® ELISA Kit?

K. Sunder

Verified customer

Asked: 2019-01-14

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may alter 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 to limit protein degradation.

Boster Scientific Support

Answered: 2019-01-14

Question

Q: we need your recommendation regarding the dilution ratio of serum samples for detection of Proteinase 3 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 low serum volumes so we like to dilute as much as possible.

M. Brown

Verified customer

Asked: 2018-07-31

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 Proteinase 3 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: 2018-07-31

Question

Q: Can Proteinase 3 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2017-10-15

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: 2017-10-15

Question

Q: how to thaw whole blood sample for Proteinase 3 ELISA after freezing?

Verified Customer

Verified customer

Asked: 2017-02-19

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 Proteinase 3 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: 2017-02-19

Question

Q: if the enzyme conjugated Proteinase 3 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 Proteinase 3 antigen?

W. Sunder

Verified customer

Asked: 2016-10-22

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: 2016-10-22

Question

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

Verified Customer

Verified customer

Asked: 2015-11-28

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: 2015-11-28

Question

Q: What is the optimal O.D. value for Proteinase 3 ELISA kit? I performed your Proteinase 3 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 Proteinase 3 even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2015-05-02

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. regarding 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 proportion 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 Proteinase 3 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Proteinase 3 that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2015-05-02

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2014-04-08

Answer

A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present 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: 2014-04-08

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