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Human Renalase (RNLS) ELISA Kit (HUEB1102)

SKU:
HUEB1102
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q5VYX0
Range:
0.78-50 ng/mL
ELISA Type:
Sandwich
Synonyms:
RNLS, Renalase, MAO-C, chromosome 10 open reading frame 59, Monoamine oxidase-C, renalase, FAD-dependent amine oxidase
Reactivity:
Human
€649
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Description

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Human Renalase (RNLS) ELISA Kit

The Human Renalase (RNLS) ELISA Kit is a highly reliable and sensitive assay designed for the precise measurement of renalase levels in human serum, plasma, and cell culture supernatants. With its high specificity and accuracy, this kit provides consistent and reproducible results, making it suitable for a variety of research applications.Renalase is an important enzyme involved in regulating blood pressure and kidney function, and its levels are known to be altered in various renal and cardiovascular diseases.

By accurately measuring renalase levels, researchers can gain valuable insights into these conditions and potentially identify new therapeutic targets.Overall, the Human Renalase (RNLS) ELISA Kit is an essential tool for studying the role of renalase in health and disease, offering researchers a reliable method for detecting and quantifying this key enzyme in biological samples.

Product Name:Human Renalase (RNLS) ELISA Kit
SKU:HUEB1102
Size:96T
Target:Human Renalase (RNLS)
Synonyms:Monoamine oxidase-C, MAO-C, C10orf59
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:0.78-50ng/mL
Sensitivity:0.32ng/mL
Intra CV:4.9%
Inter CV:8.1%
Linearity:
Sample1:21:41:81:16
Serum(N=5)89-99%81-91%99-107%105-115%
EDTA Plasma(N=5)107-117%92-102%88-98%101-110%
Heparin Plasma(N=5)106-116%101-111%87-97%101-111%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum109103-115
Plasma111105-117
Function:Catalyzes the oxidation of the less abundant 1,2-dihydro-beta-NAD(P) and 1,6-dihydro-beta-NAD(P) to form beta-NAD(P)(+). The enzyme hormone is secreted by the kidney, and circulates in blood and modulates cardiac function and systemic blood pressure. Lowers blood pressure in vivo by decreasing cardiac contractility and heart rate and preventing a compensatory increase in peripheral vascular tone, suggesting a causal link to the increased plasma catecholamine and heightened cardiovascular risk. High concentrations of catecholamines activate plasma renalase and promotes its secretion and synthesis.
Uniprot:Q5VYX0
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Renalase
Research Area:Cell Biology
Subcellular Location:Secreted
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:RENALASE: Probable FAD-dependent amine oxidase secreted by the kidney, which circulates in blood and modulates cardiac function and systemic blood pressure. Degrades catecholamines such as dopamine, norepinephrine and epinephrine in vitro. Lowers blood pressure in vivo by decreasing cardiac contractility and heart rate and preventing a compensatory increase in peripheral vascular tone, suggesting a causal link to the increased plasma catecholamine and heightened cardiovascular risk. High concentrations of catecholamines activate plasma renalase and promotes its secretion and synthesis. According to PubMed:17385068, is unlikely that renalase has physiologically relevant catecholamine-oxidizing activity. Belongs to the renalase family. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Oxidoreductase; Secreted, signal peptide; Secreted; EC 1.6.3.5

Chromosomal Location of Human Ortholog: 10q23.31

Cellular Component: extracellular space

Molecular Function:oxidoreductase activity

Biological Process: heart contraction; regulation of systemic arterial blood pressure; epinephrine metabolic process; phosphate ion homeostasis; norepinephrine metabolic process; dopamine metabolic process

NCBI Summary:Renalase is a flavin adenine dinucleotide-dependent amine oxidase that is secreted into the blood from the kidney (Xu et al., 2005 [PubMed 15841207]).[supplied by OMIM, Mar 2008]
UniProt Code:Q5VYX0
NCBI GenInfo Identifier:73914006
NCBI Gene ID:55328
NCBI Accession:Q5VYX0.1
UniProt Secondary Accession:Q5VYX0,Q9BS33, Q9NUP8,
UniProt Related Accession:Q5VYX0
Molecular Weight:34,949 Da
NCBI Full Name:Renalase
NCBI Synonym Full Names:renalase, FAD-dependent amine oxidase
NCBI Official Symbol:RNLS
NCBI Official Synonym Symbols:C10orf59; RENALASE
NCBI Protein Information:renalase; MAO-C; alpha-NAD(P)H oxidase/anomerase; monoamine oxidase-C
UniProt Protein Name:Renalase
UniProt Synonym Protein Names:Monoamine oxidase-C; MAO-C; alpha-NAD(P)H oxidase/anomerase
UniProt Gene Name:RNLS
UniProt Entry Name:RNLS_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.
ELISA
Human Renalase ELISA Kit

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