Rat EGR2 (Early Growth Response Protein 2) CLIA Kit (RTES00192)

SKU:
RTES00192
€649

Description

ELISA Kit Technical ManualMSDS

Rat EGR2 (Early Growth Response Protein 2) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat EGR2 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat EGR2 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 EGR2, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat EGR2. The concentration of Rat EGR2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range62.50-4000 pg/mL
Sensitivity37.50 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat EGR2 in samples. No significant cross-reactivity or interference between Rat EGR2 and analogues was observed.

Typical Data

As the RLU 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
(pg/mL)
RLU Average Corrected
400054978
63184
5908159046
200025540
30178
2785927824
100013505
13463
1348413449
5006427
6783
66056570
2503495
2991
32433208
1251626
1536
15811546
62.50722
788
755720
035
35
35--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat EGR2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat EGR2 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)197.35449.301828.86203.89442.921988.48
Standard deviation20.5645.96148.3224.9246.68143.37
C V (%)10.4210.238.1112.2210.547.21

Recovery

The recovery of Rat EGR2 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)89-10495
EDTA plasma (n=5)97-113103
Cell culture media (n=5)100-112106

Linearity

Samples were spiked with high concentrations of Rat EGR2 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 (%)88-10085-9791-108
Average (%)959198
1:4Range (%)85-99101-11899-116
Average (%)92109107
1:8Range (%)101-11789-10383-97
Average (%)1099590
1:16Range (%)94-10992-10393-106
Average (%)10197101

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 CLIA 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 Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Rat EGR2 (Early Growth Response Protein 2) CLIA Kit (RTES00192) 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Rat EGR2 (Early Growth Response Protein 2) CLIA Kit (RTES00192) Protein Information

UniProt Protein Function:EGR1: Transcriptional regulator. Recognizes and binds to the DNA sequence 5'-CGCCCCCGC-3'(EGR-site). Activates the transcription of target genes whose products are required for mitogenesis and differentiation. By growth factors. Belongs to the EGR C2H2-type zinc-finger protein family.
UniProt Protein Details:

Protein type:DNA-binding; C2H2-type zinc finger protein; Transcription factor

Chromosomal Location of Human Ortholog: 5q31.1

Cellular Component: nucleoplasm; cytoplasm; nucleus

Molecular Function:histone acetyltransferase binding; protein binding; DNA binding; sequence-specific DNA binding; metal ion binding; double-stranded DNA binding; transcription factor binding; transcription factor activity

Biological Process: regulation of long-term neuronal synaptic plasticity; transcription from RNA polymerase II promoter; response to nutrient levels; circadian rhythm; positive regulation of transcription, DNA-dependent; cytokine and chemokine mediated signaling pathway; response to amphetamine; negative regulation of transcription from RNA polymerase II promoter; response to cocaine; BMP signaling pathway; response to ethanol; learning and/or memory; cellular response to insulin stimulus; positive regulation of neuron apoptosis; response to glucose stimulus; oligodendrocyte differentiation; positive regulation of transcription from RNA polymerase II promoter; response to electrical stimulus; negative regulation of apoptosis; regulation of protein sumoylation; T cell differentiation

NCBI Summary:The protein encoded by this gene belongs to the EGR family of C2H2-type zinc-finger proteins. It is a nuclear protein and functions as a transcriptional regulator. The products of target genes it activates are required for differentitation and mitogenesis. Studies suggest this is a cancer suppresor gene. [provided by RefSeq, Jul 2008]
UniProt Code:P51774
NCBI GenInfo Identifier:4503493
NCBI Gene ID:1958
NCBI Accession:NP_001955.1
UniProt Secondary Accession:P51774,P11161, P08046, P08152, P08154, P51774,
UniProt Related Accession:P18146
Molecular Weight:57,507 Da
NCBI Full Name:early growth response protein 1
NCBI Synonym Full Names:early growth response 1
NCBI Official Symbol:EGR1
NCBI Official Synonym Symbols:TIS8; AT225; G0S30; NGFI-A; ZNF225; KROX-24; ZIF-268
NCBI Protein Information:early growth response protein 1; EGR-1; zinc finger protein 225; transcription factor ETR103; transcription factor Zif268; zinc finger protein Krox-24; nerve growth factor-induced protein A
UniProt Protein Name:Early growth response protein 1
UniProt Synonym Protein Names:AT225; Nerve growth factor-induced protein A; NGFI-A; Transcription factor ETR103; Transcription factor Zif268; Zinc finger protein 225; Zinc finger protein Krox-24
Protein Family:E3 SUMO-protein ligase
UniProt Gene Name:EGR1
UniProt Entry Name:EGR1_HUMAN
View AllClose

Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
CLIA
View AllClose