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Product Info Summary
| SKU: | EK1566 |
|---|---|
| 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 info
Product Name
Human Legumain(total) ELISA Kit PicoKine®
View all Legumain/Asparaginyl Endopeptidase ELISA kits
SKU/Catalog Number
EK1566
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Legumain (total) ELISA Kit PicoKine® (96 Tests). Quantitate Human LGMN in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/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
Human Legumain(total) ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1566)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: V18-Y433
Sensitivity
<10 pg/ml
Assay Range
31.2 pg/ml - 2,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1566 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1566 is reactive to LGMN in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK1566 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Legumain/Asparaginyl Endopeptidase
Legumain (asparaginyl endopeptidase, citvac, proteinase B, hemoglobinase, PRSC1 gene product or LGMN (Homo sapiens), vicilin peptidohydrolase, bean endopeptidase) is an enzyme that in humans is encoded by the LGMN gene (previous symbolPRSC1). Using fluorescence in situ hybridization, the LGMN gene was mapped to chromosome 14q32.1. This enzyme may be involved in the processing of bacterial peptides and endogenous proteins for MHC class II presentation in the lysosomal/endosomal systems. Enzyme activation is triggered by acidic pH and appears to be autocatalytic. Protein expression occurs after monocytes differentiate into dendritic cells. A fully mature, active enzyme is produced following lipopolysaccharide expression in mature dendritic cells. Overexpression of this gene may be associated with the majority of solid tumor types.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1566-CAP | Anti-Human LGMN Pre-coated 96-well strip microplate | 1 |
| EK1566-ST | Human LGMN Standard | 2 vials, 10 ng/tube |
| EK1566-DA | Human LGMN 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 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.108 | 0.179 | 0.240 | 0.298 | 0.458 | 0.755 | 1.327 | 1.928 |
Data Example Images
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Human Legumain(total) PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Legumain/Asparaginyl Endopeptidase, Dilution ratio of 1:10, concentration in serum and plasma is around 400 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Legumain(total) 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) | 86 | 299 | 1057 | 90 | 289 | 1074 |
| Standard deviation | 4.33 | 23.62 | 45.45 | 6.21 | 26.58 | 54.77 |
| CV (%) | 6.9% | 7.9% | 4.3% | 6.9% | 9.2% | 5.1% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Legumain/Asparaginyl Endopeptidase 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 | 86 | 81 | 89 | 82 | 84 | 3.2 | 3.8% |
| Sample 2 | 299 | 271 | 286 | 316 | 293 | 16.56 | 5.6% |
| Sample 3 | 1057 | 903 | 1033 | 909 | 975 | 70.04 | 7.1% |
Protein Target Info & Infographic
Gene/Protein Information For LGMN (Source: Uniprot.Org, NCBI)
Gene Name
LGMN
Full Name
Legumain
Weight
49411 MW
Superfamily
peptidase C13 family
Alternative Names
Legumain;3.4.22.34;Asparaginyl endopeptidase;Protease, cysteine 1;LGMN;PRSC1; LGMN AEP1, PRSC1, LGMN legumain legumain|asparaginyl endopeptidase|cysteine protease 1|protease, cysteine 1|protease, cysteine, 1 (legumain)
*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 LGMN, check out the LGMN 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 LGMN: 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 Human Legumain(total) ELISA Kit PicoKine® (EK1566)
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Customer Reviews
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Customer Q&As
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11 Customer Q&As for Human Legumain(total) ELISA Kit PicoKine®
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2020-10-24
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 a must for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2020-10-24
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2020-09-17
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 might differ by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2020-09-17
Question
Q: we need your recommendation regarding the dilution ratio of serum samples for detection of Legumain in Human plasma? I am trying to measure a multiple 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.
Verified Customer
Verified customer
Asked: 2020-05-19
Answer
A: without having an understanding 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 Legumain 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: 2020-05-19
Question
Q: how to proceed with the analysis of ELISA data? I have performed Legumain level in plasma.
Verified Customer
Verified customer
Asked: 2019-03-23
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: 2019-03-23
Question
Q: how to thaw whole blood sample for Legumain ELISA after freezing?
Verified Customer
Verified customer
Asked: 2019-01-31
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 Legumain 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: 2019-01-31
Question
Q: What is the optimal O.D. value for Legumain ELISA kit? I used your Legumain 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 Legumain even though the O.D. values are not very high?
E. Krishna
Verified customer
Asked: 2018-09-03
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. 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. 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 Legumain ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Legumain that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2018-09-03
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2018-07-03
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: 2018-07-03
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
W. Turner
Verified customer
Asked: 2018-03-15
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 can impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-03-15
Question
Q: if the enzyme conjugated Legumain 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 Legumain antigen?
Verified Customer
Verified customer
Asked: 2017-08-08
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: 2017-08-08
Question
Q: Can Legumain ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2016-09-13
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 suitable 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 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: 2016-09-13
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification Legumain using the EK1566 Human Legumain Picokine® ELISA Kit?
H. Adams
Verified customer
Asked: 2014-06-15
Answer
A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may change 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 repetitive freeze-thaw cycles with the stored samples to avoid protein degradation.
Boster Scientific Support
Answered: 2014-06-15
