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Cat.No.:E-EL-0060
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Assay type | Competitive-ELISA |
Format | 96T/48T |
Assay time | 2.0h |
Detection range | 31.25-2000 ng/mL |
Sensitivity | 18.75 ng/mL |
Sample type &Sample volume | serum, plasma and other biological fluids; 50μL |
Specificity | This kit recognizes MDA in samples. No significant cross-reactivity or interference between MDA 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 MDA concentrations in serum, plasma and other biological fluids. |
Item | Specifications | Storage |
---|---|---|
Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips |
-20℃, 12 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), 12 months |
Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 12 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℃ |
Plate Sealer | 5 pieces | |
Manual | 1 copy | |
Certificate of Analysis | 1 copy |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level MDA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level MDA 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) | 109.60 | 308.70 | 848.80 | 111.50 | 321.00 | 892.50 |
Standard deviation | 6.10 | 17.30 | 37.30 | 7.00 | 17.00 | 46.40 |
CV (%) | 5.57 | 5.60 | 4.39 | 6.28 | 5.30 | 5.20 |
The recovery of MDA 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) | 92-103 | 97 |
EDTA plasma(n=8) | 94-107 | 99 |
Cell culture media(n=8) | 93-110 | 100 |
Research Area | Neuroscience, Signal transduction, Metabolism |
1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C |
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2. Aspirate and wash the plate for 3 times |
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3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times |
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4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C |
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5. Add 50μL Stop Solution |
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6. Read the plate at 450nm immediately. Calculation of the results |
PMID: 33663539 |
Sample: Tissue Homogenate |
Sample: Testicular |
DOI: 10.1002/ptr.7113 |
|
PMID: 33899292 |
Sample: Serum |
Sample: Brain |
PMID: 36091371 |
Sample: Lung,Serum,Testis |
Sample: Serum |
DOI: 10.1093/ijnp/pyy090 |
|
Sample: Plasma,Tissue Homogenate |
Sample: Serum |
PMID: 36330218 |
Sample: Liver |
PMID: 32532068 |
Sample: Serum |
DOI: 10.3390/ijms22042069 |
|
PMID: 33669786 |
Sample: Tissue(Left Ventricular) |
Sample: Huvecs |
PMID: 35912092 |
Sample: Lm16 Cell,Lm36 Cell |
Sample: Plasma |
Sample: Serum |
DOI: 10.1155/2020/8078412 |
|
PMID: 32695177 |
Sample: Serum |
Sample: Serum,Sciatic Nerves |
Sample: Adrenal Tissue |
Sample: Serum |
PMID: 34266654 |
Sample: Serum |
PMID: 31824881 |
Sample: Serum,Tissue Homogenate |
Sample: Heart Tissue |
Sample: Tissue Homogenate |
Sample: Serum |
Sample: Serum |
Sample: Serum |
PMID: 27840933 |
Sample: Serum |
Sample: Plasma,Renal,Liver |
DOI: 10.1111/jfbc.13258 |
|
Sample: Serum |
Sample: Serum |
PMID: 35507975 |
Sample: Plasma,Vastus Intermedius Muscle Tissue |
PMID: 32744464 |
Sample: Tissue Homogenate |
PMID: 35331657 |
Sample: Serum,Cochlea |
PMID: 34180746 |
Sample: Ocular |
PMID: 32738910 |
Sample: Tissue Homogenate |
Sample: Lo2 Cells |
DOI: 10.3967/bes2018.101 |
|
Sample: Plasma,Tissue Homogenate |
DOI: 10.12659/MSM.924372 |
|
Sample: Kidney Tissue,Nrk-52E Cell |
Sample: Aortic Tissues |
PMID: 31892574 |
Sample: Plasma |
PMID: 32988895 |
Sample: Plasma |
PMID: 33047165 |
Sample: Serum |
PMID: 28943754 |
Sample: Lens Tissue |
PMID: 30820147 |
Sample: Lens Tissue |
DOI: 10.3390/ani10081383 |
|
PMID: 32785083 |
Sample: Plasma |
PMID: 33590347 |
Sample: Tissue Homogenate(Liver) |
DOI: 10.2147/JPR.S234334 |
|
Sample: Plasma |
Sample: Skin |
PMID: 35240291 |
Sample: Serum |
Sample: Cell Culture Supernatant |
PMID: 36219935 |
Sample: Blood |
Sample: Serum |
Sample: Tissue Homogenate |
Sample: Serum |
DOI: 10.1155/2021/5577875 |
|
Sample: Tissue Homogenate |
DOI: 10.1155/2022/2814196 |
|
PMID: 35502173 |
Sample: Plasma |
DOI: 10.1155/2021/8367802 |
|
PMID: 33643424 |
Sample: Plasma |
DOI: 10.1155/2022/7832540 |
|
Sample: Hepatic Tissue |
Sample: Kidney |
Sample: Serum |
DOI: 10.1111/jfbc.14058 |
|
PMID: 34981526 |
Sample: Serum |
Sample: Serum |
DOI: 10.1002/iub.1321 |
|
Sample: Cell Lysate |
PMID: 32489292 |
Sample: Serum |
PMID: 33076878 |
Sample: Serum |
PMID: 32803042 |
Sample: Serum |
Sample: Tissue Homogenate |
Sample: Hippocampus |
PMID: 34490475 |
Sample: Serum |
Sample: Plasma |
DOI: 10.7717/peerj.13257 |
|
PMID: 35673387 |
Sample: Sv40 Mes 13 Cell |
PMID: 34009457 |
Sample: Serum |
PMID: 34871743 |
Sample: Hippocampus,Liver |
Sample: Serum,Tissue Homogenate |
Sample: Serum |
PMID: 33002592 |
Sample: Serum |
Sample: Testicular |
PMID: 32339907 |
Sample: Serum |
PMID: 35933083 |
Sample: Serum |
Sample: Serum |
PMID: 33937367 |
Sample: Serum,Tissue Homogenate,Tissue Slice |
DOI: 10.1155/2022/9539676 |
|
PMID: 36330203 |
Sample: Plasma |
Sample: Cochlear Tissue |
Sample: Serum |
PMID: 30599959 |
Sample: Serum |
Sample: Plasma |
Sample: Serum |
DOI: 10.1002/fsn3.3096 |
|
Sample: Cerebellum,Cerebrum |
PMID: 35058758 |
Sample: Hippocampus Tissue |
PMID: 35298730 |
Sample: Hippocampal,Hepatic |
PMID: 31785243 |
Sample: Serum |
DOI: 10.1002/jbt.22483 |
|
Sample: Serum |
PMID: 36121373 |
Sample: Plasma |
Sample: Brain |
PMID: 35413439 |
Sample: Ac16 Cell |
PMID: 35552996 |
Sample: Hippocampus |
PMID: 32488017 |
Sample: Plasma,Tissue Homogenate |
Sample: Cardiac Tissue |
Sample: Testicular Tissue |
Sample: Podocytes |
PMID: 36385866 |
Sample: Serum |
DOI: 10.2147/DDDT.S291591 |
|
PMID: 33707940 |
Sample: Tissue Homogenate |
Sample: Brain |
PMID: 35029831 |
Sample: Hepatic Tissue |
PMID: 35420345 |
Sample: Serum |
PMID: 34216342 |
Sample: Serum |
DOI: 10.1111/jcmm.13569 |
|
Sample: Plasma |
DOI: 10.2147/DDDT.S344247 |
|
PMID: 35965961 |
Sample: Serum |
PMID: 34504131 |
Sample: Serum |
PMID: 34274339 |
Sample: Myocardial |
PMID: 36227296 |
Sample: Blood |
Sample: Serum |
PMID: 32497865 |
Sample: Serum |
Sample: Tissue Homogenate |
DOI: 10.3390/nu12051522 |
|
PMID: 32456230 |
Sample: Serum |
DOI: 10.3390/nu12103077 |
|
PMID: 33050310 |
Sample: Serum |
PMID: 35805580 |
Sample: Plasma |
PMID: 36009399 |
Sample: Plasma |
Sample: Serum,Liver |
Sample: Serum |
DOI: 10.3390/jpm12050833 |
|
PMID: 35629255 |
Sample: Serum |
DOI: 10.3390/md20050308 |
|
Sample: Human Dermal Fibroblasts (Hdfs) |
PMID: 34801930 |
Sample: Hffs Cells,Hsfs Cells |
PMID: 36032243 |
Sample: Serum |
PMID: 31425004 |
Sample: Serum,Cell Culture Medium |
DOI: 10.1159/000492995 |
|
Sample: Htr8/Svneo Cells |
Sample: Plasma |
PMID: 35558879 |
Sample: Hcmecs |
Sample: Arpe-19 Cell |
Sample: Lung Tissues |
DOI: 10.3390/nu13051480 |
|
PMID: 33925704 |
Sample: Serum |
DOI: 10.3390/nu13020434 |
|
PMID: 33572715 |
Sample: Serum |
PMID: 35292015 |
Sample: Serum |
Sample: Serum |
PMID: 35032726 |
Sample: Testis |
PMID: 34356359 |
Sample: Plasma |
PMID: 33671691 |
Sample: Serum |
PMID: 35671584 |
Sample: Serum |
DOI: 10.1155/2021/8889313 |
|
PMID: 33628393 |
Sample: Bronchoalveolar Lavage Fluid |
DOI: 10.1155/2021/6678924 |
|
PMID: 33574979 |
Sample: Cell Lysate |
DOI: 10.3390/nu14235025 |
|
Sample: Serum,Placental |
DOI: 10.1155/2022/7288729 |
|
PMID: 35915612 |
Sample: Bronchoalveolar Lavage Fluid (Balf),Huvecs |
DOI: 10.1155/2022/9041914 |
|
Sample: Lung Tissue |
PMID: 35417750 |
Sample: Mammary Gl,Tissue |
Sample: Serum |
PMID: 36309297 |
Sample: Seminal Plasma |
PMID: 32234613 |
Sample: Tissue Homogenate |
PMID: 35640876 |
Sample: Leaf |
Sample: Human Foreskin Fibroblasts (Hffs),Hsf Cell |
PMID: 35961822 |
Sample: Primary Kupffer Cell |
N****** Submitted [ Dec 11 2023 ]
Asked: Thank you for your response. I detected protein concentration by bradford assay resulting in concentration of (example: 61, 225, 190 mg/ml). How can I normalise protein concentration? Should it be within nanorange to be determined by the kit?
Reply
adminSubmitted [ Dec 11 2023 ]
Answered: The tissue samples were made into 10% tissue homogenate, and then the total concentration was detected by BCA, and then ELISA was performed. The concentration of target protein/total tissue protein detected by ELISA method can be obtained the concentration of target protein in the tissue sample (pg/mg prot). It is our suggestion that we give to you. If you have the calculation of other detection methods, you can also search the relevant information for reference calculation.
N****** Submitted [ Dec 11 2023 ]
Asked: After dilution and homogenization of the sample with 1:9 PBS, we need to measure the protein concentration in the sample. I am curious as to why this step is necessary.
Reply
adminSubmitted [ Dec 11 2023 ]
Answered: Hi Nourhan, We just suggest that you can choose to detect the protein concentration, you can choose according to your own experimental needs. Because at present many indicators for detecting tissue homogenates in the literature are required for protein quantification, this suggestion is to make the experimental results of the customer in the publication of the article more complete.
adminSubmitted [ Dec 11 2023 ]
Answered: Because of the different sizes of the tissue blocks and the efficiency of protein extraction, the total protein content of the tissue homogenates extracted from different samples varies within a certain range, or the protein content varies significantly between samples. These can make subsequent data processing difficult, and it is not possible to distinguish whether the variability is due to the effect of the study treatment or to the protein content. Therefore, the protein quantification of tissue homogenate is highly recommend. This ensures that data from different groups of samples are compared on the same baseline, resulting in a more accurate presentation of the data.