Human PTPRN (Protein Tyrosine Phosphatase Receptor Type N) ELISA Kit
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PTPRN . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PTPRN and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human PTPRN, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm 2 nm. The OD value is proportional to the concentration of Human PTPRN. The concentration of Human PTPRN in samples can be calculated by comparing the OD of the samples to the standard curve.
|Detection Range||0.31-20 ng/mL|
|Sample Volume Required Per Well||100uL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Human PTPRN in samples. No significant cross-reactivity or interference between Human PTPRN and analogues was observed.
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PTPRN were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PTPRN were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.38||4.89||5.39||6.74||4.49||3.70|
The recovery of Human PTPRN spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||90-100||95|
|Cell culture media (n=5)||88-100||94|
Samples were spiked with high concentrations of Human PTPRN and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
Kit Components & Storage
An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100X)||1 vial, 120 uL|
|Concentrated HRP Conjugate (100X)||1 vial, 120 uL||-20'C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4'C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25X)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4'C(shading light)|
|Stop Solution||1 vial, 10 mL||4'C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
Human PTPRN (Protein Tyrosine Phosphatase Receptor Type N) ELISA Kit (HUES02031) Assay procedure
- 1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- 2. Aliquot 100 µL of standard solutions into the standard wells.
- 3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- 4. Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- 5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- 6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37 °C.
- 7. Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- 8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- 9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 10. Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- 11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- 12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
Human PTPRN (Protein Tyrosine Phosphatase Receptor Type N) ELISA Kit (HUES02031) Protein Information
|UniProt Protein Function:||PTPRN: Implicated in neuroendocrine secretory processes. May be involved in processes specific for neurosecretory granules, such as their biogenesis, trafficking or regulated exocytosis or may have a general role in neuroendocrine functions. Seems to lack intrinsic enzyme activity. May play a role in the regulation of secretory granules via its interaction with SNTB2. Belongs to the protein-tyrosine phosphatase family. Receptor class 8 subfamily. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Membrane protein, integral; Receptor protein phosphatase, tyrosine; Motility/polarity/chemotaxis
Chromosomal Location of Human Ortholog: 2q35-q36.1
Cellular Component: integral to membrane
Molecular Function:GTPase binding; protein binding; spectrin binding; protein tyrosine phosphatase activity
Biological Process: response to reactive oxygen species; response to cAMP; response to estrogen stimulus; insulin secretion; cytokine and chemokine mediated signaling pathway; response to glucose stimulus; protein amino acid dephosphorylation; response to insulin stimulus
|NCBI Summary:||The protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. This PTP possesses an extracellular region, a single transmembrane region, and a single catalytic domain, and thus represents a receptor-type PTP. This PTP was found to be an autoantigen that is reactive with insulin-dependent diabetes mellitus (IDDM) patient sera, and thus may be a potential target of autoimmunity in diabetes mellitus. Alternate splicing results in multiple transcript variants.[provided by RefSeq, Dec 2010]|
|NCBI GenInfo Identifier:||2499754|
|NCBI Gene ID:||5798|
|UniProt Secondary Accession:||Q16849,Q08319, Q53QD6, Q6NSL1, B4DK12, F5GZS3,|
|UniProt Related Accession:||Q16849|
|NCBI Full Name:||Receptor-type tyrosine-protein phosphatase-like N|
|NCBI Synonym Full Names:||protein tyrosine phosphatase, receptor type, N|
|NCBI Official Symbol:||PTPRN|
|NCBI Official Synonym Symbols:||IA2; IA-2; ICA512; R-PTP-N; IA-2/PTP|
|NCBI Protein Information:||receptor-type tyrosine-protein phosphatase-like N; ICA 512; PTP IA-2; islet cell antigen 2; islet cell antigen 512; islet cell autoantigen 3; protein tyrosine phosphatase-like N|
|UniProt Protein Name:||Receptor-type tyrosine-protein phosphatase-like N|
|UniProt Synonym Protein Names:||Islet cell antigen 512; ICA 512; Islet cell autoantigen 3; PTP IA-2|
|Protein Family:||Receptor-type tyrosine-protein phosphatase|
|UniProt Gene Name:||PTPRN|
|UniProt Entry Name:||PTPRN_HUMAN|