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Human ALDH2/Mitochondrial Aldehyde Dehydrogenase  ELISA Kit

SKU:
HUFI02195
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P05091
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
ALDH, ALDH2, Aldehyde dehydrogenase, mitochondrial, ALDHI, ALDH class 2, ALDH-E2, ALDM, Aldehyde Dehydrogenase 2
Reactivity:
Human
€599
Frequently bought together:

Description

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Human ALDH2/Mitochondrial Aldehyde Dehydrogenase ELISA Kit

The Human ALDH2 (Mitochondrial Aldehyde Dehydrogenase) ELISA Kit is a powerful tool for accurately measuring ALDH2 levels in human samples including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures reliable and reproducible results, making it suitable for a wide range of research applications.ALDH2, also known as Mitochondrial Aldehyde Dehydrogenase, is an important enzyme involved in the oxidation of aldehydes, playing a crucial role in alcohol metabolism and detoxification processes.

Variations in ALDH2 activity have been linked to various diseases, including cardiovascular diseases, cancer, and neurodegenerative disorders, making it a valuable biomarker for research and drug development in these areas.Overall, the Human ALDH2 ELISA Kit provides researchers with a reliable and efficient method for studying ALDH2 levels and its implications in human health and disease.

Product Name:Human ALDH2 / Mitochondrial Aldehyde Dehydrogenase  ELISA Kit
Product Code:HUFI02195
Size:96 Assays
Alias:ALDH, ALDH2, Aldehyde dehydrogenase, mitochondrial, ALDHI, ALDH class 2, ALDH-E2, ALDM, Aldehyde Dehydrogenase 2
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human ALDH concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human ALDH and the recovery rates were calculated by comparing the measured value to the expected amount of Human ALDH in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10090
EDTA plasma(n=5)86-10494
UFH plasma(n=5)87-10096
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human ALDH and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)92-100%86-93%85-102%
EDTA plasma(n=5)88-100%84-101%83-93%
UFH plasma(n=5)84-91%83-100%80-92%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP05091
UniProt Protein Function:ALDH2: This protein belongs to the aldehyde dehydrogenase family of proteins. Aldehyde dehydrogenase is the second enzyme of the major oxidative pathway of alcohol metabolism. Two major liver isoforms of aldehyde dehydrogenase, cytosolic and mitochondrial, can be distinguished by their electrophoretic mobilities, kinetic properties, and subcellular localizations. Most Caucasians have two major isozymes, while approximately 50% of Orientals have the cytosolic isozyme but not the mitochondrial isozyme. A remarkably higher frequency of acute alcohol intoxication among Orientals than among Caucasians could be related to the absence of a catalytically active form of the mitochondrial isozyme. The increased exposure to acetaldehyde in individuals with the catalytically inactive form may also confer greater susceptibility to many types of cancer. This gene encodes a mitochondrial isoform, which has a low Km for acetaldehydes, and is localized in mitochondrial matrix. Alternative splicing results in multiple transcript variants encoding distinct isoforms.[provided by RefSeq, Mar 2011]
UniProt Protein Details:Protein type:Lipid Metabolism - glycerolipid; Lipid Metabolism - fatty acid; Secondary Metabolites Metabolism - limonene and pinene degradation; Amino Acid Metabolism - valine, leucine and isoleucine degradation; EC 1.2.1.3; Amino Acid Metabolism - histidine; Carbohydrate Metabolism - ascorbate and aldarate; Mitochondrial; Carbohydrate Metabolism - propanoate; Oxidoreductase; Amino Acid Metabolism - arginine and proline; Carbohydrate Metabolism - pyruvate; Amino Acid Metabolism - tryptophan; Carbohydrate Metabolism - glycolysis and gluconeogenesis; Other Amino Acids Metabolism - beta-alanine; Amino Acid Metabolism - lysine degradation; Carbohydrate Metabolism - butanoate

Chromosomal Location of Human Ortholog: 12q24.2

Cellular Component: mitochondrial matrix

Molecular Function:aldehyde dehydrogenase (NAD) activity; electron carrier activity; aldehyde dehydrogenase [NAD(P)+] activity

Biological Process: synaptic transmission; ethanol catabolic process; xenobiotic metabolic process; alcohol metabolic process; carbohydrate metabolic process; ethanol oxidation; neurotransmitter biosynthetic process

Disease: Alcohol Sensitivity, Acute

NCBI Summary:This protein belongs to the aldehyde dehydrogenase family of proteins. Aldehyde dehydrogenase is the second enzyme of the major oxidative pathway of alcohol metabolism. Two major liver isoforms of aldehyde dehydrogenase, cytosolic and mitochondrial, can be distinguished by their electrophoretic mobilities, kinetic properties, and subcellular localizations. Most Caucasians have two major isozymes, while approximately 50% of East Asians have the cytosolic isozyme but not the mitochondrial isozyme. A remarkably higher frequency of acute alcohol intoxication among East Asians than among Caucasians could be related to the absence of a catalytically active form of the mitochondrial isozyme. The increased exposure to acetaldehyde in individuals with the catalytically inactive form may also confer greater susceptibility to many types of cancer. This gene encodes a mitochondrial isoform, which has a low Km for acetaldehydes, and is localized in mitochondrial matrix. Alternative splicing results in multiple transcript variants encoding distinct isoforms.[provided by RefSeq, Nov 2016]
UniProt Code:P05091
NCBI GenInfo Identifier:118504
NCBI Gene ID:217
NCBI Accession:P05091.2
UniProt Related Accession:P05091
Molecular Weight:
NCBI Full Name:Aldehyde dehydrogenase, mitochondrial
NCBI Synonym Full Names:aldehyde dehydrogenase 2 family member
NCBI Official Symbol:ALDH2  
NCBI Official Synonym Symbols:ALDM; ALDHI; ALDH-E2  
NCBI Protein Information:aldehyde dehydrogenase, mitochondrial
UniProt Protein Name:Aldehyde dehydrogenase, mitochondrial
UniProt Synonym Protein Names:ALDH class 2; ALDH-E2; ALDHI
Protein Family:NADP-dependent fatty aldehyde dehydrogenase
UniProt Gene Name:ALDH2  
UniProt Entry Name:ALDH2_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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