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Product Info Summary
| SKU: | EK1765 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA, citrate). |
Product info
Product Name
Human PAM ELISA Kit PicoKine®
View all Peptidylglycine alpha-Amidating Monooxygenase/PAM ELISA kits
SKU/Catalog Number
EK1765
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human PAM ELISA Kit PicoKine™ (96 Tests). Quantitate Human PAM in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 75pg/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 PAM ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1765)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: NS0; Immunogen sequence: M1-V817
Sensitivity
<75 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1765 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1765 is reactive to PAM in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
Application Guarantee
EK1765 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Peptidylglycine alpha-Amidating Monooxygenase/PAM
Peptidyl-glycine alpha-amidating monooxygenase is an enzyme that catalyzes the biosynthesis of many signaling peptides and, in humans, is encoded by the PAM gene. It is mapped to 5q21.1. This gene encodes a multifunctional protein. The encoded preproprotein is proteolytically processed to generate the mature enzyme. This enzyme includes two domains with distinct catalytic activities, a peptidylglycine alpha-hydroxylating monooxygenase (PHM) domain and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain. These catalytic domains work sequentially to catalyze the conversion of neuroendocrine peptides to active alpha-amidated products. Alternative splicing results in multiple transcript variants, at least one of which encodes an isoform that is proteolytically processed.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1765-CAP | Anti-Human PAM Pre-coated 96-well strip microplate | 1 |
| EK1765-ST | Human PAM Standard | 2 vials, 10 ng/tube |
| EK1765-DA | Human PAM 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 | 156 | 312 | 625 | 1,250 | 2,500 | 5,000 | 10,000 |
| O.D. | 0.028 | 0.054 | 0.161 | 0.387 | 0.732 | 1.161 | 1.457 | 2.093 |
Data Example Images
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Human PAM PicoKine ELISA Kit Standard Curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Peptidylglycine alpha-Amidating Monooxygenase/PAM, Dilution ratio of 1:50, concentration in serum is 21.7-36.9 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human PAM 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) | 164 | 572 | 4574 | 250 | 468 | 3375 |
| Standard deviation | 4.1 | 31.17 | 116.18 | 11.65 | 34.77 | 146.48 |
| CV(%) | 2.5% | 5.5% | 2.5% | 4.7% | 7.4% | 4.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Peptidylglycine alpha-Amidating Monooxygenase/PAM 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 | 164 | 228 | 291 | 184 | 217 | 4.51 | 2.1% |
| Sample 2 | 572 | 532 | 446 | 618 | 542 | 39.73 | 7.3% |
| Sample 3 | 4574 | 4967 | 4068 | 4322 | 4483 | 147.48 | 3.3% |
Protein Target Info & Infographic
Gene/Protein Information For PAM (Source: Uniprot.Org, NCBI)
Gene Name
PAM
Full Name
Peptidyl-glycine alpha-amidating monooxygenase
Weight
Alternative Names
PAL; PAM; pancreatic peptidylglycine alpha-amidating monooxygenase; peptidyl alpha-amidating enzyme; peptidyl-alpha-hydroxyglycine alpha-amidating lyase; peptidylamidoglycolate lyase; peptidylglycine 2-hydroxylase; peptidyl-glycine alpha-amidating monooxygenase; peptidylglycine alpha-amidating monooxygenase; peptidylglycine alpha-hydroxylating monooxygenase; PHM PAM PAL, PHM peptidylglycine alpha-amidating monooxygenase peptidyl-glycine alpha-amidating monooxygenase|pancreatic peptidylglycine alpha-amidating monooxygenase|peptidyl alpha-amidating enzyme|peptidyl-alpha-hydroxyglycine alpha-amidating lyase|peptidylamidoglycolate lyase|peptidylglycine 2-hydroxylase|peptidylglycine alpha-hydroxylating monooxygenase
*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 PAM, check out the PAM 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 PAM: 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 PAM ELISA Kit PicoKine® (EK1765)
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Customer Q&As
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7 Customer Q&As for Human PAM ELISA Kit PicoKine®
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of PAM in Human plasma? I am trying to measure a multiple 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: 2020-08-24
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 PAM 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: 2020-08-24
Question
Q: if the enzyme conjugated PAM 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 PAM antigen?
R. Gonzalez
Verified customer
Asked: 2019-09-05
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: 2019-09-05
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
T. Adams
Verified customer
Asked: 2019-05-29
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: 2019-05-29
Question
Q: how to proceed with the analysis of ELISA data? I have obtained PAM level in plasma.
G. Baker
Verified customer
Asked: 2019-04-19
Answer
A: please read 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: 2019-04-19
Question
Q: What is the optimal O.D. value for PAM ELISA kit? I performed your PAM 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 PAM even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2018-01-27
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 protocol, 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 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 PAM ELISA kit to have sensitivity of 50pg/ml, that means the minimum amount of PAM that can be declared/interpreted as positive by the above standard is 50pg/ml.
Boster Scientific Support
Answered: 2018-01-27
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
H. Lopez
Verified customer
Asked: 2017-07-29
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 needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2017-07-29
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring PAM using the EK1765 Human PAM Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2016-01-27
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 right after collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples to avoid protein degradation.
Boster Scientific Support
Answered: 2016-01-27
