Sandwich ELISA to human Sterol O-acyltransferase 1 AEE0003

$560,00$800,00

ELISA kit for human Sterol O-acyltransferase 1, with data demonstrating no cross-reactivity to other CoA-acyltransferases.

Description

ACAT1 / SOAT1

All names and symbols: Acyl-coenzyme A:cholesterol acyltransferase 1; Cholesterol acyltransferase 1; Sterol O-acyltransferase 1; ACAT; ACACT; ACAT1; ACAT-1; SOAT; STAT;

Confirmed species reactivityHuman
Sensitivity4.75 pg/ml
Linear calculation range15.625 – 500 pg/ml
Precision intra-assay / inter-assay<10% / <12%
Aeonian Rating®90
RRID
MatrixRecovery Range (%)Average (%)
Serum (n=5)80-10291
EDTA plasma (n=5)81-10090
Heparin plasma (n=5)80-8984
Recovery: Matrices were spiked with recombinant ACAT1 and the recovery rates were calculated by comparing the measured value to the expected amount of ACAT1 in the matrices.
Matrix1:21:41:81:16
Serum (n=5)87-91%87-107%74-101%92-97%
EDTA plasma (n=5)90-105%84-101%90-101%79-108%
Heparin plasma (n=5)84-95%92-105%82-105%89-91%
Linearity: The linearity of the kit was assayed by testing serially diluted matrices spiked with a fixed amount of recombinant ACAT1. The results were demonstrated by the percentage of calculated concentration related to the expected concentration of ACAT1 in the diluted matrices.

Calibratrion curve:

AEE0003 to ACAT1 was successfully used to determine the concentration of ACAT1 in serum, plasma and buffer. Recommended serum dilution 1:16, and EDTA plasma dilution 1:8.

Calibration curve of recombinant ACAT1 in buffer. HRP detection by Absorbance at 450nm.

Selectivity of the ELISA kit:

AEE0003 to ACAT1 was successfully confirmed selective as there were no signals found in the presence of other CoA-acyltransferases at high and low concentrations.

Selectivity was tested by comparing the signals of recombinant ACAT1 with related other recombinant CoA-acyltranferases, each at two concentrations. The other CoA-acyltranferases tested were ACAT2, and OXCT1. HRP detection by Absorbance at 450nm.

ELISA Test:

The microtiter plate provided in this kit has been pre-coated with an antibody specific to ACAT1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to ACAT1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain ACAT1, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ACAT1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

ELISA Procedure:

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37℃;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
8. Add 50µL Stop Solution. Read at 450nm immediately.

Stability:

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions.
Note:
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

Product Manual Traditional
Product Manual Ready to Use
Material Safety Data Sheet Traditional
Material Safety Data Sheet Ready to Use

Additional information

DL-ACAT1-Hu (traditional) 96T

Composition:
Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2
Standard 2
Diluents buffer 1×45ml
Detection Reagent A 1×120μl
Detection Reagent B 1×120μl
TMB Substrate 1×9ml
Stop Solution 1×6ml
Wash Buffer (30 × concentrate) 1×20ml
Instruction manual 1

DL-ACAT1-Hu (traditional) 48T

Composition:
Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2
Standard 2
Diluents buffer 1×22.5ml
Detection Reagent A 1×60μl
Detection Reagent B 1×60μl
TMB Substrate 1×4.5ml
Stop Solution 1×3ml
Wash Buffer (30 × concentrate) 1×10ml
Instruction manual 1

DLR-ACAT1-Hu (ready-to-use) 96T

Composition:
Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2
Standard 2
Standard Diluent 1×20ml
Detection Solution A 1×12ml
Detection Solution B 1×12ml
TMB Substrate 1×9ml
Stop Solution 1×6ml
Wash Buffer (30 × concentrate) 1×20ml
Instruction manual 1

DLR-ACAT1-Hu (ready-to-use) 48T

Composition:
Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2
Standard 2
Standard Diluent 1×10ml
Detection Solution A 1×6ml
Detection Solution B 1×6ml
TMB Substrate 1×4.5ml
Stop Solution 1×3ml
Wash Buffer (30 × concentrate) 1×10ml
Instruction manual 1

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SKU: AEE0003 Category: