null

Rat Adenosine A3 Receptor / ADORA3 ELISA Kit (RTFI00555)

SKU:
RTFI00555
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P28647
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
ADORA3
Reactivity:
Rat
€649
Frequently bought together:

Description

system_update_altDatasheetsystem_update_altMSDS

Rat Adenosine A3 Receptor/ADORA3 ELISA Kit

The Rat Adenosine A3 Receptor (ADORa3) ELISA Kit is specifically designed for the quantitative detection of ADORa3 levels in rat serum, plasma, and tissue homogenates. This kit offers high precision and accuracy, ensuring consistent and reliable results for various research applications.ADORa3 is a key receptor involved in regulating various physiological processes, including inflammation, neurotransmission, and immune response. Dysregulation of ADORa3 signaling has been associated with conditions such as chronic pain, neurological disorders, and inflammatory diseases, making it a valuable target for therapeutic interventions.

With its high sensitivity and specificity, the Rat ADORa3 ELISA Kit is an essential tool for researchers studying the role of ADORa3 in disease pathogenesis and exploring potential treatment strategies. Order now to advance your research in the field of adenosine receptor biology.

Product Name:Rat ADORA3 (Adenosine A3 Receptor) ELISA Kit
Product Code:RTFI00555
Size:96 Assays
Target:Rat ADORA3
Alias:ADORA3
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat ADORA3 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat ADORA3 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10597
EDTA plasma(n=5)85-10297
UFH plasma(n=5)90-9592
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat ADORA3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)87-105%89-100%91-105%
EDTA plasma(n=5)87-96%83-100%84-101%
UFH plasma(n=5)90-97%87-95%81-100%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P28647
UniProt Protein Function:ADORA3: Receptor for adenosine. The activity of this receptor is mediated by G proteins which inhibits adenylyl cyclase. Possible role in reproduction. Belongs to the G-protein coupled receptor 1 family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:GPCR, family 1; Membrane protein, multi-pass; Receptor, GPCR; Membrane protein, integral

Chromosomal Location of Human Ortholog: 1p13.2

Cellular Component: integral to membrane; plasma membrane

Molecular Function:adenosine receptor activity, G-protein coupled

Biological Process: adenosine receptor signaling pathway

NCBI Summary:This gene encodes a protein that belongs to the family of adenosine receptors, which are G-protein-coupled receptors that are involved in a variety of intracellular signaling pathways and physiological functions. The receptor encoded by this gene mediates a sustained cardioprotective function during cardiac ischemia, it is involved in the inhibition of neutrophil degranulation in neutrophil-mediated tissue injury, it has been implicated in both neuroprotective and neurodegenerative effects, and it may also mediate both cell proliferation and cell death. Alternative splicing results in multiple transcript variants. This gene shares its 5' terminal exon with some transcripts from overlapping GeneID:57413, which encodes an immunoglobulin domain-containing protein. [provided by RefSeq, Nov 2014]
UniProt Code:P28647
NCBI GenInfo Identifier:1351831
NCBI Gene ID:140
NCBI Accession:P33765.2
UniProt Secondary Accession:P28647,Q61618, P28647,
UniProt Related Accession:P0DMS8
Molecular Weight:
NCBI Full Name:
NCBI Synonym Full Names:adenosine A3 receptor
NCBI Official Symbol:ADORA3  
NCBI Official Synonym Symbols:A3AR  
NCBI Protein Information:adenosine receptor A3
UniProt Protein Name:
Protein Family:Adenosine receptor
UniProt Gene Name:
StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37°C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

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 clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum 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 anti-coagulant. 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 & homogenizein 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.

Related Products