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Human LDHA(Lactate Dehydrogenase A) ELISA Kit

Citations(8)Uniprot : P00338

Cat.No.:E-EL-H0556
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.78-50 ng/mL
Sensitivity	0.47 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human LDHA in samples. No significant cross-reactivity or interference between Human LDHA 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 Human LDHA 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 Human LDHA. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human LDHA 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 Human LDHA, 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 Human LDHA. You can calculate the concentration of Human LDHA 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 Human LDHA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human LDHA 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)	2.30	7.50	20.90	2.20	8.20	21.30
Standard deviation	0.10	0.30	0.90	0.10	0.40	1.10
CV (%)	4.35	4.00	4.31	4.55	4.88	5.16

Recovery

The recovery of Human LDHA 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)	96-109	101
EDTA plasma (n=8)	86-97	92
Cell culture media (n=8)	91-105	97

Linearity

Samples were spiked with high concentrations of Human LDHA and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P00338
Synonyms	GSD11, LDH1, LDHM
Research Area	Cancer, Metabolism, Signal transduction

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

American Journal of Translational Research (2021) IF: 4.06
The effect of coenzyme Q10 plus trimetazidine on acute viral myocarditis treatment

PMID: 35035725
Sample: serum
BONE (2020) IF: 4.147
Inhibition of glycolysis by targeting lactate dehydrogenase A facilitates hyaluronan synthase 2 synthesis in synovial fibroblasts of temporomandibular joint osteoarthritis

DOI: 10.1016/j.bone.2020.115584

Sample: synovial fluid
Toxics (2023) IF: 4.472
A Proposed Association between Improving Energy Metabolism of HepG2 Cells by Plant Extracts and Increasing Their Sensitivity to Doxorubicin

DOI: 10.3390/toxics11020182

Sample: HepG2 cell
TOXICOLOGY (2022) IF: 4.571
Evaluation of mutagenic, cytotoxic, mitochondrial dysfunction, apoptotic activity, and acute toxicity of ethanolic extract of Cissus quadrangularis

DOI: 10.1016/j.tox.2022.153373

Sample: HepG2 cell
Bioengineered (2021) IF: 3.269
MiR-181a-5p facilitates proliferation, invasion, and glycolysis of breast cancer through NDRG2-mediated activation of PTEN/AKT pathway

DOI: 10.1080/21655979.2021.2006974

PMID: 34951340
Sample: MDA-MB-231 cell,SK-BR-3 cell
Annals of Translational Medicine (2021) IF: 3.932
Combination of oncolytic adenovirus ZD55 harboring TRAIL-IETD-MnSOD and cytokine-induced killer cells against lung cancer

DOI: 10.21037/atm-21-4479

PMID: 34790733
Sample: serum,BEAS-2B cell
CYTOTECHNOLOGY (2021) IF: 2.058
Investigation of glucose catabolism in hypoxic Mcf 7 breast cancer culture

DOI: 10.1007/s10616-021-00459-2

PMID: 33927477
Sample: Cell（MCF-7 cancer cells）
International Journal of Medical Sciences (2020) IF: 2.523
Downregulation of P300/CBP-Associated Factor Attenuates Myocardial Ischemia-Reperfusion Injury Via Inhibiting Autophagy

DOI: 10.7150/ijms.44604

PMID: 32547315
Sample: Serum,Cell culture supernatant

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