HMOX1 Kit ELISA

N° du produit ABIN455320

Détail du produit HMOX1 Kit ELISA

Antigène: HMOX1 (Heme Oxygenase (Decycling) 1 (HMOX1))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 0.78-50 ng/mL

Seuil minimal de détection: 0.78 ng/mL

Application: ELISA

Fonction: This immunoassay kit allows for the specific measurement of human HO-1 concentrations in serum, and plasma.

Type d'échantillon: Serum, Plasma

Analytical Method: Quantitative

Specificité: This assay recognizes recombinant and natural human HO-1.

Réactivité croisée (Details): No significant cross-reactivity or interference was observed.

Attributs du produit: Homo sapiens,Human,Heme oxygenase 1,HO-1,HMOX1,HO,HO1,1.14.99.3

Ingrédients: Reagent (Quantity): Assay plate (1), Standard (2), Sample Diluent (1x20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A 1×120µl Detection Reagent B 1×120µl Wash Buffer(25 x concentrate) (1x30ml), Substrate (1x10ml), Stop Solution (1x10ml)

Information d'application

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for HO-1 has been pre-coated onto a microplate. Standards and samples are 2 pipetted into the wells and any HO-1 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for HO-1 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of HO-1 bound in the initial step. The color development is stopped and the intensity of the color is measured.

Préparation des réactifs:

Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix 3 gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 500 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 50 ng/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (50 ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL). Detection Reagent A and B - Dilute to the working concentration specified on the vial label using Assay Diluent A and B (1:100), respectively.

Prélèvement de l'échantillon: Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 - 8 °C within 30 minutes of collection. Store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Note: Citrate plasma has not been validated for use in this assay.

Procédure de l'essai:

Allow all reagents to reach room temperature. Arrange and label required number of strips.
1. Prepare all reagents, working standards and samples as directed in the previous sections.
2. Add 100 uL of Standard, Control, or sample per well. Cover with the adhesive strip. Incubate for 2 hours at 37 °C.
3. Remove the liquid of each well, don’t wash.
4. Add 100 uL of Detection Reagent A to each well. Incubate for 1 hour at 37°C. Detection Reagent A may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
5. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (350 uL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
6. Add 100 uL of Detection Reagent B to each well. Cover with a new adhesive strip.Incubate for 1 hours at 37 °C.
7. Repeat the aspiration/wash as in step
5. 8. Add 90 uL of Substrate Solution to each well. Incubate for 30 minutes at room temperature. Protect from light.
9. Add 50 uL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
10. Determine the optical density of each well within 30 minutes, using a microplate reader set to 450 nm.
Important Note:
1. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
2. It is recommended that no more than 32 wells be used for each assay run if manual pipetting is used since pipetting of all standards, specimens and controls should be completed within 5 minutes. A full plate of 96 wells may be used if automated pipetting is available.
3. Duplication of all standards and specimens, although not required, is recommended. 4
4. When mixing or reconstituting protein solutions, always avoid foaming.
5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
6. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

Calcul des résultats:

Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the HO-1 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: 1. The kit should not be used beyond the expiration date on the kit label.
2. Do not mix or substitute reagents with those from other lots or sources.
3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

Stock: 4 °C/-20 °C

Stockage commentaire: The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.

Détail du antigène

Antigène: HMOX1 (Heme Oxygenase (Decycling) 1 (HMOX1))

Autre désignation: HMOX1 (HMOX1 Produits)

Synonymes: HMOX1 Kit ELISA, ARABIDOPSIS THALIANA HEME OXYGENASE 1 Kit ELISA, ATHO1 Kit ELISA, F18A8.4 Kit ELISA, F18A8_4 Kit ELISA, GENOMES UNCOUPLED 2 Kit ELISA, GUN2 Kit ELISA, HEME OXYGENASE Kit ELISA, HEME OXYGENASE 1 Kit ELISA, HEME OXYGENASE 6 Kit ELISA, HO1 Kit ELISA, HY1 Kit ELISA, HY6 Kit ELISA, PLASTID HEME OXYGENASE Kit ELISA, REVERSAL OF THE DET PHENOTYPE 4 Kit ELISA, HO-1 Kit ELISA, wu:fc27c04 Kit ELISA, zgc:65984 Kit ELISA, D8Wsu38e Kit ELISA, Hemox Kit ELISA, Hmox Kit ELISA, Hsp32 Kit ELISA, HEOXG Kit ELISA, Heox Kit ELISA, Ho-1 Kit ELISA, Ho1 Kit ELISA, hsp32 Kit ELISA, MGC132176 Kit ELISA, Hmox1 Kit ELISA, HMOX1D Kit ELISA, HSP32 Kit ELISA, bK286B10 Kit ELISA, heme oxygenase 1 Kit ELISA, Plant heme oxygenase (decyclizing) family protein Kit ELISA, Heme oxygenase Kit ELISA, heme oxygenase Kit ELISA, heme oxygenase 1, chloroplastic Kit ELISA, heme oxygenase 1a Kit ELISA, heme oxygenase 1 L homeolog Kit ELISA, HMOX1 Kit ELISA, TED4 Kit ELISA, ho1 Kit ELISA, P9301_RS17555 Kit ELISA, A9601_RS17590 Kit ELISA, MAE_RS06565 Kit ELISA, HO1 Kit ELISA, hmox1a Kit ELISA, CPE0214 Kit ELISA, Cyan7425_2962 Kit ELISA, Hmox1 Kit ELISA, hmox1.L Kit ELISA, AM1_C0205 Kit ELISA, CC1G_11686 Kit ELISA, CpipJ_CPIJ007246 Kit ELISA

Sujet: Heme Oxygenase-1 (HO-1) also known as Hsp32, is the inducible isoform of heme oxygenase that catalyzes the NADPH, O2 and cytochrome P450 reductase dependent oxidation of heme to carbon monoxide, ferrous iron and biliverdin which is rapidly reduced to bilirubin. These products of the HO reaction have important physiological effects: carbon monoxide is a potent vasodilator and has been implicated to be a physiological regulator of cGMP and vascular tone, biliverdin and its product bilirubin are potent antioxidants, “free” iron increases oxidative stress and regulates the expression of many mRNAs (e.g., DCT-1, ferritin and transferring receptor) by affecting the conformation of iron regulatory protein (IRP)-1 and its binding to iron regulatory elements (IREs) in the 5’- or 3’- UTRs of the mRNAs. To date, three identified heme oxygenase isoforms are part of the HO system that catalyze heme into biliverdin and carbon monoxide. These are inducible HO-1 or Hsp32, constitutive HO-2 that is abundant in the brain and testis, and HO-3 which is related to HO-2 but is the product of a different gene. The HO system is the rate-limiting step in heme degradation and HO activity decreases the levels of heme which is a well known potent catalyst of lipid peroxidation and oxygen radical formation. The expression of HO-1 is highly responsive to all types of stimuli that cause oxidative stress and it is up regulated during exposure to oxidants, UV-A irradiation and a series of agents including cytokines, hormones, heme and heavy metals. HO-1 is a vital component of neuronal defense mechanisms and oxidative stress has been postulated to be the underlying basis for neuronal cell death in neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease. The expression of HO-1 is normally very low in the brain but increases markedly after heat shock, ischemia or glutathione depletion. Spatial distribution of HO-1 expression in AD brain is essentially identical to that of the pathogenic conformational changes of tau protein that is the major component of the intraneuronal lesion of AD, neurofibrillary tangles.

Pathways: Transition Metal Ion Homeostasis, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, SARS-CoV-2 Protein Interactome