Rat c-myc (c-myc Oncogene Product) ELISA Kit (RTES00831)

SKU:
RTES00831
€649

Description

ELISA Kit Technical ManualMSDS

Rat c-myc (c-myc Oncogene Product) 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 c-myc . 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 c-myc 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 c-myc, 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 c-myc. The concentration of Rat c-myc 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 Range0.16-10 ng/mL
Sensitivity0.10 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Rat c-myc in samples. No significant cross-reactivity or interference between Rat c-myc 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
102.509
2.523
2.5162.445
51.691
1.711
1.7011.63
2.50.978
0.962
0.970.899
1.250.481
0.515
0.4980.427
0.630.276
0.272
0.2740.203
0.320.182
0.164
0.1730.102
0.160.117
0.129
0.1230.052
00.061
0.081
0.071--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.460.854.400.480.814.57
Standard deviation0.030.040.130.030.030.20
C V (%)6.524.712.956.253.704.38

Recovery

The recovery of Rat c-myc 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)94-110102
EDTA plasma (n=5)95-109102
Cell culture media (n=5)89-10095

Linearity

Samples were spiked with high concentrations of Rat c-myc 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 (%)90-10486-9789-104
Average (%)959196
1:4Range (%)91-10680-9382-94
Average (%)988589
1:8Range (%)87-10185-10089-99
Average (%)949194
1:16Range (%)90-10681-9386-98
Average (%)968692

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 c-myc (c-myc Oncogene Product) ELISA Kit (RTES00831) 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 c-myc (c-myc Oncogene Product) ELISA Kit (RTES00831) Protein Information

UniProt Protein Function:Myc: a proto-oncogenic transcription factor that plays a role in cell proliferation, apoptosis and in the development of human tumors. Seems to activate the transcription of growth-related genes.
UniProt Protein Details:

Protein type:Oncoprotein; Transcription factor; DNA-binding; Nucleolus

Cellular Component: axon; cytoplasm; mitochondrion; nuclear body; nucleolus; nucleoplasm; nucleus; perinuclear region of cytoplasm; protein complex; spindle

Molecular Function:DNA binding; double-stranded DNA binding; protein binding; protein complex binding; protein dimerization activity; protein heterodimerization activity; sequence-specific DNA binding; transcription factor activity; transcription factor binding; ubiquitin protein ligase binding

Biological Process: amino acid transport; B cell apoptosis; caspase activation; cell cycle arrest; cell proliferation; cellular iron ion homeostasis; cellular response to insulin stimulus; chromatin remodeling; chromosome organization and biogenesis; detection of mechanical stimulus involved in sensory perception of sound; DNA damage response, signal transduction resulting in induction of apoptosis; flavonoid metabolic process; G0 to G1 transition; glucose metabolic process; hypothalamus development; in utero embryonic development; inner mitochondrial membrane organization and biogenesis; lactic acid secretion; MAPKKK cascade; middle ear morphogenesis; negative regulation of cell division; negative regulation of fibroblast proliferation; negative regulation of glucose import; negative regulation of monocyte differentiation; negative regulation of protein binding; negative regulation of transcription from RNA polymerase II promoter; ovarian follicle development; ovulation; pathogenesis; pigmentation; positive regulation of B cell apoptosis; positive regulation of caspase activity; positive regulation of catalytic activity; positive regulation of cell cycle; positive regulation of cell proliferation; positive regulation of DNA binding; positive regulation of epithelial cell proliferation; positive regulation of fibroblast proliferation; positive regulation of glycolysis; positive regulation of mesenchymal cell proliferation; positive regulation of smooth muscle cell migration; positive regulation of smooth muscle cell proliferation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; protein processing; pyruvate transport; re-entry into mitotic cell cycle; regulation of apoptosis; regulation of gene expression; regulation of mitotic cell cycle; regulation of oxidative phosphorylation; regulation of telomere maintenance; regulation of transcription, DNA-dependent; response to alkaloid; response to DNA damage stimulus; response to drug; response to estradiol stimulus; response to ethanol; response to gamma radiation; response to radiation; response to wounding; skeletal morphogenesis; transcription from RNA polymerase II promoter; transcription initiation; transcription, DNA-dependent; transformation of host cell by virus; ureteric bud branching; Wnt receptor signaling pathway; Wnt receptor signaling pathway through beta-catenin

NCBI Summary:The protein encoded by this gene is a multifunctional, nuclear phosphoprotein that plays a role in cell cycle progression, apoptosis and cellular transformation. It functions as a transcription factor that regulates transcription of specific target genes. Mutations, overexpression, rearrangement and translocation of this gene have been associated with a variety of hematopoietic tumors, leukemias and lymphomas, including Burkitt lymphoma, in human. There is evidence to show that alternative translation initiations from an upstream, in-frame non-AUG (CUG) and a downstream AUG start site result in the production of two isoforms with distinct N-termini, in human and mouse. Rat mRNA also has a similarly placed CUG upstream of the AUG start site, suggesting that it may also produce two Myc proteins. [provided by RefSeq, Jul 2008]
UniProt Code:P09416
NCBI GenInfo Identifier:71834866
NCBI Gene ID:24577
NCBI Accession:NP_036735.2
UniProt Secondary Accession:P09416,Q6B500,
UniProt Related Accession:P09416
Molecular Weight:48,898 Da
NCBI Full Name:myc proto-oncogene protein
NCBI Synonym Full Names:myelocytomatosis oncogene
NCBI Official Symbol:Myc  
NCBI Official Synonym Symbols:mMyc; c-myc; RNCMYC  
NCBI Protein Information:myc proto-oncogene protein
UniProt Protein Name:Myc proto-oncogene protein
UniProt Synonym Protein Names:Proto-oncogene c-Myc; Transcription factor p64
Protein Family:Myc protein
UniProt Gene Name:Myc  
UniProt Entry Name:MYC_RAT
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Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
Sandwich
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