Mouse TRPV1(Transient Receptor Potential Cation Channel Subfamily V, Member 1)ELISA Kit (MOES01265)

(No reviews yet) Write a Review
SKU:
MOES01265
€549

Description

ELISA Kit Technical ManualMSDS

Mouse TRPV1(Transient Receptor Potential Cation Channel Subfamily V, Member 1)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 Mouse TRPV1 . 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 Mouse TRPV1 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 Mouse TRPV1, 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 Mouse TRPV1. The concentration of Mouse TRPV1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
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 Mouse TRPV1 in samples. No significant cross-reactivity or interference between Mouse TRPV1 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
202.508
2.554
2.5312.443
101.615
1.615
1.6151.527
50.969
0.965
0.9670.879
2.50.484
0.51
0.4970.409
1.250.308
0.288
0.2980.21
0.630.197
0.183
0.190.102
0.310.132
0.148
0.140.052
00.084
0.092
0.088--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse TRPV1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse TRPV1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.942.888.130.982.748.09
Standard deviation0.060.120.310.070.160.39
C V (%)6.384.173.817.145.844.82

Recovery

The recovery of Mouse TRPV1 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)85-9991
EDTA plasma (n=5)88-9893
Cell culture media (n=5)88-10495

Linearity

Samples were spiked with high concentrations of Mouse TRPV1 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 (%)97-11297-11185-95
Average (%)10310290
1:4Range (%)89-10580-9282-96
Average (%)968788
1:8Range (%)87-9985-9787-99
Average (%)939294
1:16Range (%)95-10687-9783-99
Average (%)1009290

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

Mouse TRPV1(Transient Receptor Potential Cation Channel Subfamily V, Member 1)ELISA Kit (MOES01265) 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.

Mouse TRPV1(Transient Receptor Potential Cation Channel Subfamily V, Member 1)ELISA Kit (MOES01265) Protein Information

UniProt Protein Function:Ligand-activated non-selective calcium permeant cation channel involved in detection of noxious chemical and thermal stimuli (PubMed:15194687, PubMed:15489017). Seems to mediate proton influx and may be involved in intracellular acidosis in nociceptive neurons. Involved in mediation of inflammatory pain and hyperalgesia (PubMed:10764638). Sensitized by a phosphatidylinositol second messenger system activated by receptor tyrosine kinases, which involves PKC isozymes and PCL. Activation by vanilloids, like capsaicin, and temperatures higher than 42 degrees Celsius, exhibits a time- and Ca2+-dependent outward rectification, followed by a long-lasting refractory state. Mild extracellular acidic pH (6.5) potentiates channel activation by noxious heat and vanilloids, whereas acidic conditions (pH <6) directly activate the channel. Can be activated by endogenous compounds, including 12-hydroperoxytetraenoic acid and bradykinin. Acts as ionotropic endocannabinoid receptor with central neuromodulatory effects. Triggers a form of long-term depression (TRPV1-LTD) mediated by the endocannabinoid anandamine in the hippocampus and nucleus accumbens by affecting AMPA receptors endocytosis.
UniProt Code:Q704Y3
NCBI GenInfo Identifier:47825364
NCBI Gene ID:193034
NCBI Accession:NP_001001445.1
UniProt Secondary Accession:Q704Y3,Q5SSE1, Q5SSE2, Q5SSE4, Q5WPV5, Q68SW0,
UniProt Related Accession:Q704Y3
Molecular Weight:93,941 Da
NCBI Full Name:transient receptor potential cation channel subfamily V member 1
NCBI Synonym Full Names:transient receptor potential cation channel, subfamily V, member 1
NCBI Official Symbol:Trpv1  
NCBI Official Synonym Symbols:Vr1; VR-1; OTRPC1; TRPV1beta; TRPV1alpha  
NCBI Protein Information:transient receptor potential cation channel subfamily V member 1
UniProt Protein Name:Transient receptor potential cation channel subfamily V member 1
UniProt Synonym Protein Names:Osm-9-like TRP channel 1; OTRPC1; Vanilloid receptor 1
Protein Family:Transient receptor potential cation channel subfamily
UniProt Gene Name:Trpv1  
View AllClose

Additional Information

Reactivity:
Mouse
ELISA Type:
Sandwich
View AllClose