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Human RNASET2 (Ribonuclease T2) ELISA Kit (HUES02424)

SKU:
HUES02424
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
O00584
Sensitivity:
0.19ng/mL
Range:
0.31-20ng/mL
ELISA Type:
Sandwich
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Immunology
€599
Frequently bought together:

Description

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Human RNASET2 (Ribonuclease T2) ELISA Kit

The Human RNASET2 (Ribonuclease T2) ELISA Kit is a reliable tool for the precise measurement of RNASET2 levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures accurate and reproducible results, making it suitable for a variety of research applications.RNASET2 is an important enzyme involved in RNA metabolism and regulation, playing a role in processes such as RNA degradation and turnover.

Dysregulation of RNASET2 has been linked to various diseases, including neurodegenerative disorders and certain cancers, making it a valuable biomarker for studying these conditions and potentially developing targeted therapies.Overall, the Human RNASET2 ELISA Kit provides researchers with a valuable tool for investigating the role of RNASET2 in disease pathology and developing new insights into potential therapeutic strategies.

Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Human
Detection Method:Colormetric
Detection Range:0.31-20 ng/mL
Sensitivity:0.19 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Human RNASET2 in samples. No significant cross-reactivity or interference between Human RNASET2 and analogues was observed.

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 RNASET2. 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 RNASET2 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 RNASET2, 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 RNASET2. The concentration of Human RNASET2 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:RNASET2: Has ribonuclease activity, with higher activity at acidic pH. May play a role in cellular RNA catabolism. Defects in RNASET2 are the cause of leukoencephalopathy cystic without megalencephaly (LCWM). An infantile- onset syndrome of cerebral leukoencephalopathy. Affected newborns develop microcephaly and neurologic abnormalities including psychomotor impairment, seizures and sensorineural hearing impairment. The brain shows multifocal white matter lesions, anterior temporal lobe subcortical cysts, pericystic abnormal myelination, ventriculomegaly and intracranial calcifications. Belongs to the RNase T2 family. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Secreted; Ribonuclease; EC 3. 1. 27. -; Secreted, signal peptide; RNA-binding

Chromosomal Location of Human Ortholog: 6q27

Cellular Component: endoplasmic reticulum lumen; extracellular region; extracellular space; lysosome

Molecular Function:ribonuclease activity

Biological Process: RNA catabolic process

Disease: Leukoencephalopathy, Cystic, Without Megalencephaly

NCBI Summary:This ribonuclease gene is a novel member of the Rh/T2/S-glycoprotein class of extracellular ribonucleases. It is a single copy gene that maps to 6q27, a region associated with human malignancies and chromosomal rearrangement. [provided by RefSeq, Jul 2008]
UniProt Code:O00584
NCBI GenInfo Identifier:20139363
NCBI Gene ID:8635
NCBI Accession:O00584. 2
UniProt Secondary Accession:O00584,Q5T8Q0, Q8TCU2, Q9BZ46, Q9BZ47, B2RDA7, E1P5C3
UniProt Related Accession:O00584
Molecular Weight:14,000 Da
NCBI Full Name:Ribonuclease T2
NCBI Synonym Full Names:ribonuclease T2
NCBI Official Symbol:RNASET2
NCBI Official Synonym Symbols:RNASE6PL; bA514O12. 3
NCBI Protein Information:ribonuclease T2
UniProt Protein Name:Ribonuclease T2
UniProt Synonym Protein Names:Ribonuclease 6
Protein Family:Ribonuclease
UniProt Gene Name:RNASET2
UniProt Entry Name:RNT2_HUMAN

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.DAverageCorrected
202.484
2.496		2.492.425
101.584
1.63		1.6071.542
50.932
0.924		0.9280.863
2.50.432
0.47		0.4510.386
1.250.267
0.255		0.2610.196
0.630.18
0.156		0.1680.103
0.310.11
0.124		0.1170.052
00.056
0.074		0.065--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human RNASET2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human RNASET2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)1.112.056.711.182.216.25
Standard deviation0.060.090.220.060.120.34
C V (%)5.414.393.285.085.435.44

Recovery

The recovery of Human RNASET2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)90-10596
EDTA plasma (n=5)86-10293
Cell culture media (n=5)87-9792

Linearity

Samples were spiked with high concentrations of Human RNASET2 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 (%)84-9995-11088-100
Average (%)9010394
1:4Range (%)88-10281-9584-97
Average (%)958788
1:8Range (%)91-10586-10084-98
Average (%)999191
1:16Range (%)94-10783-9688-104
Average (%)998895

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 ×12 strips -20°C, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 µL
Concentrated HRP Conjugate (100×) 1 vial, 120 µL -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 (25×) 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
  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 100µl of standard solutions into the standard wells.
  3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
  4. Add 100µl 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 100µL 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 350µL 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 100µL 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 90µL 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.

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