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A-CoA(Acetyl Coenzyme A) ELISA Kit

Cat.No.:E-EL-0125
Reactivity: Universal

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes A-CoA in samples. No significant cross-reactivity or interference between A-CoA and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of A-CoA concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Universal A-CoA. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Universal A-CoA and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Universal A-CoA, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of 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 Universal A-CoA. You can calculate the concentration of Universal A-CoA in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level A-CoA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level A-CoA were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.11	2.68	7.94	1.00	2.48	7.46
Standard deviation	0.08	0.12	0.40	0.07	0.11	0.27
CV (%)	7.21	4.48	5.04	7.00	4.44	3.62

Recovery

The recovery of A-CoA 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=8)	88-99	94
EDTA plasma (n=8)	86-97	91
Cell culture media (n=8)	93-108	99

Target Information

Research Area

Tags and Cell Markers, Cardiovascular, Metabolism

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

BIOMATERIALS (2022) IF: 15.304
Self-amplified ROS production from fatty acid oxidation enhanced tumor immunotherapy by atorvastatin/PD-L1 siRNA lipopeptide nanoplexes

DOI: 10.1016/j.biomaterials.2022.121902

Sample: B16-F10 cell,CT-26 cell
EMBO JOURNAL (2022) IF: 14.012
CRIP1 suppresses BBOX1-mediated carnitine metabolism to promote stemness in hepatocellular carcinoma

DOI: 10.15252/embj.2021110218

PMID: 35775648
Sample: Huh7 cell,Hep3B cell,MHCC-97H cell,HepG2 cell
Journal of Integrative Plant Biology (2022) IF: 7.061
Vitamin B1 THIAMIN REQUIRING1 synthase mediates the maintenance of chloroplast function by regulating sugar and fatty acid metabolism in rice

DOI: 10.1111/jipb.13283

PMID: 35603832
Sample: leaf,seedling
BIOCHEMICAL PHARMACOLOGY (2022) IF: 6.1
Lamivudine remedies alcoholism by activating acetaldehyde dehydrogenase

DOI: 10.1016/j.bcp.2022.115199

PMID: 35917871
Sample: liver
Pharmaceutics (2022) IF: 6.525
Two-in-One Nanoparticle Formulation to Deliver a Tyrosine Kinase Inhibitor and microRNA for Targeting Metabolic Reprogramming and Mitochondrial Dysfunction in Gastric Cancer

DOI: 10.3390/pharmaceutics14091759

PMID: 36145507
Sample: AGS cell
LIFE SCIENCES (2023) IF: 6.78
Alfuzosin ameliorates diabetes by boosting PGK1 activity in diabetic mice

DOI: 10.1016/j.lfs.2023.121491

Sample: Liver tissue
Oxidative Medicine and Cellular Longevity (2020) IF: 5.076
Mild Hypothermia Attenuates Hepatic Ischemia–Reperfusion Injury through Regulating the JAK2/STAT3-CPT1a-Dependent Fatty Acid β-Oxidation

DOI: 10.1155/2020/5849794

Sample: Tissue homogenate
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS (2023) IF: 4.117
Sulfathiazole treats type 2 diabetes by restoring metabolism through activating CYP19A1

DOI: 10.1016/j.bbagen.2023.130303

Sample: liver tissue
MOLECULAR AND CELLULAR ENDOCRINOLOGY (2018) IF: 3.563
Curcumin restrains hepatic glucose production by blocking cAMP/PKA signaling and reducing acetyl CoA accumulation in high-fat diet (HFD)-fed mice

DOI: 10.1016/j.mce.2018.02.018

Sample: liver,primary hepatocytes
Journal of Diabetes and Metabolic Disorders (2022)
Primaquine activates Keratin 7 to treat diabetes and its complications

DOI: 10.1007/s40200-022-01135-8

PMID: 36404863
Sample: plasma

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Reviews/Q&A

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Reviews
Q&A

W********hSubmitted [ Mar 07 2023 ]

Asked: hi. I am a researcher working at the institute for basic science, South Korea. We purchased your kit (i.e. Acetyl CoA elisa kit) this time and are trying to utilize it, but strangely, there is a reaction with your reference/standard sample, but there is no reaction with the Acetyl CoA we have (Roche and Sigma-aldrich, two products). We are wondering if your product is actually capable of accurately measuring acetyl CoA and would like to get some advice from your technical team. We look forward to your response. Best regards

adminSubmitted [ Mar 07 2023 ]

Answered: Dear Wuhyun Koh, Thanks for your feedback. At present, we can guarantee the applicability and detection rate of the E-EL-0125 for natural samples only. We have verified a large number of serum, plasma and some tissue samples, and the detection effect conforms to the literature report very well. We have not done a lot of validation on some recombinant proteins and other products so far, and there may indeed be undetectable situations. We have searched Merck's official website for A-CoA products. Sigma Aldrich contains A-CoA-3Na (Cat No.: A2056) and A-CoA-Li+(Cat No.: A2181), and Roche products (SKU: 10101893001; 10101907001; 11585371001) contain A-CoA-Li3 according to the chemical composition description on the product page. We speculate that there may be some difference between those product above and composition of our standard in A-CoA ELISA kit, which lead to the the possibility of undetected. A-coa is a chemical (Acetyl-CoA C23H38N7O17P3S), and it's a small molecule, which is not applicable the spatial structure/label commonly used for conventional large molecules In addition, different manufacturers have different methods for A-CoA synthesis, and the reagents or other factors added in the process of may have some influence on reaction system of ELISA, thus affecting the detected OD value. Based the above situation, we recommend you to use natural samples to detect. Thanks for your kind understanding. If any further questions, pls feel free to contact customer00@elabscience.com Best regards, Crystal

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