Rat GALR1 (Galanin Receptor1) ELISA Kit (RTES00982)

SKU:
RTES00982
Product type:
ELISA
Reactivity:
Rat
ELISA Type:
Sandwich
€649

Description

ELISA Kit Technical ManualMSDS

Rat GALR1 (Galanin Receptor1) 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 Rat GALR1 . 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 Rat GALR1 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 Rat GALR1, 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 Rat GALR1. The concentration of Rat GALR1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection MethodColormetric
Detection Range0.78-50 ng/mL
Sensitivity0.47 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Rat GALR1 in samples. No significant cross-reactivity or interference between Rat GALR1 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
502.49
2.53
2.512.446
251.677
1.695
1.6861.622
12.50.988
0.954
0.9710.907
6.250.437
0.439
0.4380.374
3.130.243
0.215
0.2290.165
1.570.173
0.159
0.1660.102
0.780.11
0.122
0.1160.052
00.058
0.07
0.064--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat GALR1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GALR1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)2.286.0622.852.206.1721.39
Standard deviation0.150.280.820.130.261.04
C V (%)6.584.623.595.914.214.86

Recovery

The recovery of Rat GALR1 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)89-10296
EDTA plasma (n=5)92-110100
Cell culture media (n=5)91-10498

Linearity

Samples were spiked with high concentrations of Rat GALR1 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 (%)96-11191-10388-103
Average (%)1029796
1:4Range (%)92-10881-9287-104
Average (%)998695
1:8Range (%)91-10382-9685-95
Average (%)978890
1:16Range (%)87-10382-9685-98
Average (%)948890

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

Rat GALR1 (Galanin Receptor1) ELISA Kit (RTES00982) 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 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.

Rat GALR1 (Galanin Receptor1) ELISA Kit (RTES00982) Protein Information

UniProt Protein Function:GALR1: Receptor for the hormone galanin. The activity of this receptor is mediated by G proteins that inhibit adenylate cyclase activity. Belongs to the G-protein coupled receptor 1 family.
UniProt Protein Details:

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

Cellular Component: integral to membrane; integral to plasma membrane; plasma membrane

Molecular Function:galanin receptor activity; neuropeptide binding; peptide hormone binding

Biological Process: elevation of cytosolic calcium ion concentration; G-protein coupled receptor protein signaling pathway; G-protein signaling, adenylate cyclase activating pathway; G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); negative regulation of adenylate cyclase activity; neuropeptide signaling pathway; positive regulation of cortisol secretion; positive regulation of transcription from RNA polymerase II promoter

NCBI Summary:G-protein coupled receptor that binds the neuropeptide galanin; inhibits cAMP formation [RGD, Feb 2006]
UniProt Code:Q62805
NCBI GenInfo Identifier:164518935
NCBI Gene ID:50577
NCBI Accession:NP_037090.2
UniProt Related Accession:Q62805
Molecular Weight:38,862 Da
NCBI Full Name:galanin receptor type 1
NCBI Synonym Full Names:galanin receptor 1
NCBI Official Symbol:Galr1  
NCBI Official Synonym Symbols:Galnr1  
NCBI Protein Information:galanin receptor type 1
UniProt Protein Name:Galanin receptor type 1
Protein Family:Galanin receptor
UniProt Gene Name:Galr1  
UniProt Entry Name:GALR1_RAT
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