CUSABIO's product CSB-E04489h is a ready-to-use ELISA kit designed for the quantitative measurement of ACE2 concentrations in serum, plasma, or cell culture supernates. It provides a detection range of 0.156 ng/mL-10 ng/mL. It has high sensitivity as low as 0.039 ng/mL. Validated parameters of this kit also include precision, linearity, and recovery.
This kit contains all reagents necessary for quantitatively detecting ACE2 levels. In this assay, an anti-human ACE2 antibody has been pre-coated on the microplate. Samples and standards are respectively added to wells. After thorough washing, a biotin-conjugated ACE2 antibody is added to wells. ACE2 in the samples or in the standards is sandwiched between the twp antibody pairs. Following a rigorous washing, HRP-avidin conjugate is pipetted into wells. The solution turns blue after the addition of the TMB substrate. The color reaction is terminated when adding the stop solution, and the solution turns blue to yellow. The color intensity is positively correlated to ACE2 concentrations in the sample and is measured by reading the absorption value at 450 nm in a microplate reader.
ACE2, on the one hand, catalyzes the conversion of Ang I and Ang II into the inactive Ang 1-9 and Ang 1-7 as a carboxypeptidase in the renin-angiotensin-aldosterone system (RAAS). Through the breakout of Ang II into other molecules, ACE2 regulates blood pressure, maintains blood pressure homeostasis, and reduces inflammation and tissue injury. On the other hand, ACE2 has been found to serve as a receptor for SARS-CoV, NL63 coronavirus, and SARS-CoV-2. In the SARS-CoV and SARS-CoV-2 infection, ACE2 on the host cells binds to viral spike (S) protein promoting the viral entry into host cells.
||angiotensin I converting enzyme (peptidyl-dipeptidase A) 2
||ACE 2 ELISA Kit; ACE related carboxypeptidase ELISA Kit; ACE-related carboxypeptidase ELISA Kit; ACE2 ELISA Kit; ACE2_HUMAN ELISA Kit; ACEH ELISA Kit; Angiotensin converting enzyme 2 ELISA Kit; Angiotensin converting enzyme homolog ELISA Kit; Angiotensin converting enzyme like protein ELISA Kit; Angiotensin I Converting Enzyme (peptidyl dipeptidase A) 2 ELISA Kit; Angiotensin I converting enzyme 2 ELISA Kit; Angiotensin-converting enzyme homolog ELISA Kit; DKFZP434A014 ELISA Kit; EC 3.4.17 ELISA Kit; metalloprotease MPROT 15 ELISA Kit; Metalloprotease MPROT15 ELISA Kit; OTTHUMP00000022963 ELISA Kit; Processed angiotensin-converting enzyme 2 ELISA Kit
||Homo sapiens (Human)
||serum, plasma, cell culture supernates
||0.156 ng/mL-10 ng/mL
|Intra-assay Precision (Precision within an assay): CV%<8%|| || || |
|Three samples of known concentration were tested twenty times on one plate to assess. || |
|Inter-assay Precision (Precision between assays): CV%<10%|| || || |
|Three samples of known concentration were tested in twenty assays to assess. || || |
| || || || || || || |
|To assess the linearity of the assay, samples were spiked with high concentrations of human ACE2 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.|
| ||Sample||Serum(n=4)|| |
|1:1||Average %||87|| |
|Range %||80-99|| |
|1:2||Average %||97|| |
|Range %||90-105|| |
|1:4||Average %||95|| |
|Range %||91-107|| |
|1:8||Average %||93|| |
|Range %||80-98|| |
|The recovery of human ACE2 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.|
|Sample Type||Average % Recovery||Range|| |
|Serum (n=5) ||92||88-98|| |
|EDTA plasma (n=4)||97||90-105|| |
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|These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|10||2.216 ||2.182 ||2.199 ||1.955 || |
|5||2.012 ||1.916 ||1.964 ||1.720 || |
|2.5||1.754 ||1.643 ||1.699 ||1.455 || |
|1.25||1.326 ||1.290 ||1.308 ||1.064 || |
|0.625||1.008 ||0.944 ||0.976 ||0.732 || |
|0.312||0.714 ||0.676 ||0.695 ||0.451 || |
|0.156||0.416 ||0.392 ||0.404 ||0.160 || |
|0||0.249 ||0.239 ||0.244 || || |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Human AE2 antibody.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled ACE2 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle) ---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubating.
- An instruction manual
|Materials not provided
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Test tubes for dilution
|ELISA Data Analysis
||Watch ELISA data processing video & download Curve Expert if needed
|ELISA kit FAQs
||Store at 2-8°C. Please refer to protocol.
||3-5 working days