Human PTPRN (Protein Tyrosine Phosphatase Receptor Type N) ELISA Kit (HUES02031)

SKU:
HUES02031
€539

Description

ELISA Kit Technical ManualMSDS

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.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range0.31-20 ng/mL
Sensitivity0.19 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human PTPRN in samples. No significant cross-reactivity or interference between Human PTPRN and analogues was observed.

Typical Data

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.

Concentration
(ng/mL)
O.D Average Corrected
202.355
2.411
2.3832.326
101.581
1.601
1.5911.534
50.89
0.87
0.880.823
2.50.385
0.413
0.3990.342
1.250.225
0.215
0.220.163
0.630.157
0.153
0.1550.098
0.310.104
0.11
0.1070.05
00.05
0.064
0.057--

Precision

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
Sample123123
n202020202020
Mean (ng/mL)0.943.076.680.893.346.75
Standard deviation0.060.150.360.060.150.25
C V (%)6.384.895.396.744.493.70

Recovery

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 (%)
Serum (n=5)94-107100
EDTA plasma (n=5)90-10095
Cell culture media (n=5)88-10094

Linearity

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)
1:2Range (%)89-10490-10493-105
Average (%)969698
1:4Range (%)89-10387-10083-97
Average (%)959389
1:8Range (%)87-10386-9985-97
Average (%)949292
1:16Range (%)92-10984-9683-97
Average (%)1009089

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.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 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 Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 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]
UniProt Code:Q16849
NCBI GenInfo Identifier:2499754
NCBI Gene ID:5798
NCBI Accession:Q16849.1
UniProt Secondary Accession:Q16849,Q08319, Q53QD6, Q6NSL1, B4DK12, F5GZS3,
UniProt Related Accession:Q16849
Molecular Weight:979
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
View AllClose

Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
View AllClose