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Human ATP5A ELISA Kit (A310691)

1 Citation
Standard Curve - Human ATP5A ELISA Kit (A310691) - Antibodies.com
90 Minute ELISA
$545
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$40
Dispatched from St. Louis, MO.
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Name
Human ATP5A ELISA Kit
Description
Human ATP5A ELISA Kit is a 90 minute sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human ATP5A in serum, plasma, and other biological fluids.
Assay Type
Sandwich (quantitative)
Principle of Assay
Human ATP5A ELISA Kit (A310691) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human ATP5A in serum, plasma or other biological fluids. An antibody specific for ATP5A has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the ATP5A present in each sample is bound to the wells by the immobilized antibody. Biotinylated Anti-ATP5A Antibody, which also binds the ATP5A present in each sample, and Streptavidin-HRP, which binds the Biotinylated Anti-ATP5A Antibody, are added and the microtiter plate is incubated. Following incubation, unbound Biotinylated Anti-ATP5A Antibody and unbound Streptavidin-HRP are removed by washing, and two substrate solutions are added to the wells. Color develops in proportion to the amount of ATP5A captured in each well. The color development is stopped by addition of stop solution which changes the color from blue to yellow and the intensity of the color is then measured. The concentration of ATP5A in the samples can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
Platform
Pre-coated Microplate (12 x 8 well strips)
Detection Type
Colorimetric
Instrument
Colorimetric Microplate Reader
Sample Type
Serum, plasma or other biological fluids.
Sensitivity
3.67 ng/L
Product Range
50-800 ng/L
Assay Time
1h 30m
Reactivity
Human
Precision
Intra-assay CV: 5.8%, Inter-assay CV: < 10%
Storage
Store at +4°C. Do not use past expiration date!
Synonyms
ATP synthase alpha chain, mitochondrial, ATP synthase subunit alpha, ATP synthase subunit alpha mitochondrial, ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle, ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, 1, ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 1, cardiac muscle, ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 2, non-cardiac muscle-like 2, ATP sythase (F1 ATPase) alpha subunit, Atp5a1, ATP5AL2, ATPA_HUMAN, ATPM, Epididymis secretory sperm binding protein Li 123m, hATP1, HEL-S-123m, MC5DN4, mitochondrial, Mitochondrial ATP synthetase, Mitochondrial ATP synthetase oligomycin resistant, Modifier of Min 2 mouse homolog, Modifier of Min 2, mouse, homolog of, MOM2, OMR, ORM, OTTHUMP00000163475
Product Note
Information online is representative. Always refer to the Product Manual inside the kit.
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Kit Components
Item Quantity Storage
Pre-Coated 96 Well Microplate 12 x 8 Well Strips +4°C
Standard Solution 500µl +4°C
Standard Diluent 3ml +4°C
Biotinylated Detection Antibody 1ml +4°C
Streptavidin-HRP 6ml +4°C
Wash Buffer (25X) 20ml +4°C
Substrate Solution A 6ml +4°C
Substrate Solution B 6ml +4°C
Stop Solution 6ml +4°C
Plate Sealers 5 Adhesive Strips -
Foil Pouch 1 Zip-Sealed Pouch -
Required Materials
The following materials are not included in the kit, but are required to run this assay:
  • Microplate reader capable of measuring absorbance at 450nm ± 10nm.
  • Squirt bottle, multichannel pipette reservoir, or automated microplate washer.
  • Graph paper or computer software capable of generating logarithmic functions.
  • Test tubes and Eppendorf tubes to prepare standards and samples.
  • Multi- and single-channel pipettes and sterile pipette tips.
  • Deionized or distilled water.
  • Absorbent paper towels.
  • 37°C incubator.
Important Notes
  • Samples cannot be diluted for use with this kit. Owing to the material used to prepare the kit, sample matrix interference may falsely depress the specificity and accuracy of the assay.
  • Sample concentrations should be estimated before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their experiments.
  • Samples containing Sodium Azide (NaN3) cannot be used in this assay. NaN3 inhibits the activity of Horseradish Peroxidase (HRP) which is used in the detection step.
  • This ELISA kit is intended for the analysis of biological samples and has not been validated for purified/recombinant proteins, industrial, or commercial materials. Performance with such samples cannot be guaranteed, as differences in expression, sequence, tags, folding, post-translational modifications, purity, activity, or buffer composition may affect assay results. For further guidance, please contact our technical support team.
Bring all reagents and samples to room temperature before use.
We recommend that all standards, controls, and samples be assayed in duplicate.
Step 1
Prepare all reagents, samples, and working standards as directed in the Product Manual.
Step 2
Determine the number of microplate strips required for the assay and insert them into the plate frame. Unused strips should be stored in the foil pouch at +4°C.
Step 3
Add 50μl of standard to the standard wells.
Step 4
Add 40μl of samples to the sample wells.
Step 5
Add 10μl of Biotinylated Detection Antibody to sample wells (but not to the standard wells).
Step 6
Add 50μl of Streptavidin-HRP to sample wells and standard wells (but not to the blank control well). Mix well. Cover with the adhesive plate sealer provided. Incubate for 60 minutes at 37°C.
Step 7
Remove the plate sealer and wash the plate 5 times with Wash Buffer. Soak wells with 300μl Wash Buffer for 30 seconds to 1 minute for each wash. For automated washing, aspirate or decant each well and wash 5 times with Wash Buffer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Blot the plate onto paper towels or other absorbent material.
Step 8
Add 50μl of Substrate Solution A to each well and then add 50μl of Substrate Solution B to each well. Cover with a new adhesive plate sealer and incubate for 10 minutes at 37°C in the dark.
Step 9
Add 50μl of Stop Solution to each well. The color in the wells should change from blue to yellow immediately. Note: If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.
Step 10
Determine the optical density (O.D. value) of each well within 10 minutes, using a microplate reader set to 450 nm.

  • Scientific Validation DataValidation Data (1)

Standard Curve - Human ATP5A ELISA Kit (A310691) - Antibodies.comEnlarge Image

Standard Curve - Human ATP5A ELISA Kit (A310691)

Representative standard curve for Human ATP5A ELISA Kit (A310691).

  • Product Citations (1)

SARS-CoV-2-Derived RNA Fragment Induces Myocardial Dysfunction via siRNA-like Suppression of Mitochondrial ATP Synthase
This publication cites Human ATP5A ELISA Kit (A310691).
Article Abstract

Myocardial injury is a critical determinant of prognosis in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection; however, its underlying mechanisms remain incompletely understood. In this study, we examined the effects of SARS-CoV-2-derived RNA fragments on human cardiomyocytes. We identified a 19-nucleotide sequence within the viral genome that shares complete sequence homology with the human F1F0 ATP synthase subunit alpha gene (ATP5A). This sequence was found to associate with Argonaute 2 (AGO2) and downregulate ATP5A expression via a mechanism analogous to RNA interference. Consequently, oxidative phosphorylation was suppressed in cardiomyocytes, leading to impaired myocardial maturation and the emergence of heart failure-like phenotypes. Notably, exosome-mimetic liposomal delivery of this RNA fragment to cardiomyocytes reproduced the ATP5A-suppressive effect. These findings suggest that SARS-CoV-2-derived RNA fragments may contribute to myocardial injury through the siRNA-like modulation of mitochondrial gene expression. Further validation in animal models and patient-derived materials is warranted.

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