Human LTA4H (Leukotriene A4 Hydrolase) ELISA Kit (HUES02444)
- SKU:
- HUES02444
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P09960
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Metabolism
Description
Human LTA4H (Leukotriene A4 Hydrolase) ELISA Kit
The Human LTA4H (Leukotriene A4 Hydrolase) ELISA Kit is specially designed to accurately measure levels of LTA4H in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers reliable and consistent results, making it an excellent choice for a variety of research purposes.LTA4H is a key enzyme involved in the synthesis of leukotrienes, which are lipid mediators involved in inflammatory responses.
Dysregulation of LTA4H has been linked to various inflammatory diseases, making it a valuable biomarker for studying these conditions and potential therapeutic interventions.Overall, the Human LTA4H ELISA Kit offers researchers a powerful tool for investigating the role of LTA4H in inflammation and exploring its potential as a target for drug development.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.31-20 ng/mL |
| Sensitivity: | 0.19 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human LTA4H in samples. No significant cross-reactivity or interference between Human LTA4H and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human LTA4H. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human LTA4H and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human LTA4H, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding 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 LTA4H. The concentration of Human LTA4H in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | LTA4H: Epoxide hydrolase that catalyzes the final step in the biosynthesis of the proinflammatory mediator leukotriene B4. Has also aminopeptidase activity. Belongs to the peptidase M1 family. 3 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:EC 3. 3. 2. 6; Hydrolase; Lipid Metabolism - arachidonic acid Chromosomal Location of Human Ortholog: 12q22 Cellular Component: nucleoplasm; cytoplasm; cytosol Molecular Function:peptidase activity; leukotriene-A4 hydrolase activity; metallopeptidase activity; zinc ion binding; epoxide hydrolase activity; aminopeptidase activity Biological Process: leukotriene biosynthetic process; peptide catabolic process; arachidonic acid metabolic process; leukotriene metabolic process; proteolysis; inflammatory response |
| NCBI Summary: | The protein encoded by this gene is an enzyme that contains both hydrolase and aminopeptidase activities. The hydrolase activity is used in the final step of the biosynthesis of leukotriene B4, a proinflammatory mediator. The aminopeptidase activity has been shown to degrade proline-glycine-proline (PGP), a neutrophil chemoattractant and biomarker for chronic obstructive pulmonary disease (COPD). Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2015] |
| UniProt Code: | P09960 |
| NCBI GenInfo Identifier: | 126353 |
| NCBI Gene ID: | 4048 |
| NCBI Accession: | P09960. 2 |
| UniProt Related Accession: | P09960 |
| Molecular Weight: | ~ 69kDa |
| NCBI Full Name: | Leukotriene A-4 hydrolase |
| NCBI Synonym Full Names: | leukotriene A4 hydrolase |
| NCBI Official Symbol: | LTA4H |
| NCBI Protein Information: | leukotriene A-4 hydrolase |
| UniProt Protein Name: | Leukotriene A-4 hydrolase |
| UniProt Synonym Protein Names: | Leukotriene A(4) hydrolase |
| Protein Family: | Leukotriene A-4 hydrolase |
| UniProt Gene Name: | LTA4H |
| UniProt Entry Name: | LKHA4_HUMAN |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 20 | 2.295 2.309 | 2.302 | 2.23 |
| 10 | 1.581 1.603 | 1.592 | 1.52 |
| 5 | 0.888 0.854 | 0.871 | 0.799 |
| 2.5 | 0.405 0.437 | 0.421 | 0.349 |
| 1.25 | 0.256 0.252 | 0.254 | 0.182 |
| 0.63 | 0.183 0.159 | 0.171 | 0.099 |
| 0.31 | 0.113 0.131 | 0.122 | 0.05 |
| 0 | 0.065 0.079 | 0.072 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human LTA4H were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human LTA4H 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) | 0.92 | 1.77 | 9.81 | 0.83 | 1.93 | 10.04 |
| Standard deviation | 0.06 | 0.10 | 0.37 | 0.05 | 0.11 | 0.45 |
| C V (%) | 6.52 | 5.65 | 3.77 | 6.02 | 5.70 | 4.48 |
Recovery
The recovery of Human LTA4H 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=5) | 90-104 | 97 |
| EDTA plasma (n=5) | 86-96 | 91 |
| Cell culture media (n=5) | 88-102 | 95 |
Linearity
Samples were spiked with high concentrations of Human LTA4H and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 93-109 | 90-102 | 93-107 |
| Average (%) | 101 | 96 | 98 | |
| 1:4 | Range (%) | 89-104 | 86-96 | 83-95 |
| Average (%) | 95 | 91 | 88 | |
| 1:8 | Range (%) | 90-101 | 86-99 | 89-103 |
| Average (%) | 96 | 91 | 95 | |
| 1:16 | Range (%) | 92-105 | 85-97 | 88-100 |
| Average (%) | 98 | 90 | 94 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°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 | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
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