SOD1 Kit ELISA

N° du produit ABIN579227

Détail du produit SOD1 Kit ELISA

Antigène: SOD1 (Superoxide Dismutase 1, Soluble (SOD1))

Reactivité: Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 1.56-100 U/mL

Seuil minimal de détection: 1.56 U/mL

Application: ELISA

Fonction: This immunoassay kit allows for the specific measurement of Rat Superoxide Dismutases (SOD) concentrations in cell culture supernates, serum, and plasma.

Type d'échantillon: Cell Culture Supernatant, Serum, Plasma

Analytical Method: Quantitative

Specificité: This assay recognizes recombinant and natural Rat SOD.

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

Attributs du produit: Rattus norvegicus,Rat,Superoxide dismutase [Cu-Zn],Sod1,1.15.1.1

Ingrédients: Reagent (Quantity ): Assay plate (1), Standard (2), Sample Diluent (1 × 20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A (1 × 120μl), Detection Reagent B (1 × 120μl), Wash Buffer (25 x concentrate) (1 × 30ml), Substrate (1x10ml), Stop Solution (1 x 10ml)

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 SOD has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any SOD present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for SOD is added to the wells. Following a wash to 2 remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of SOD 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. 3 Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix 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 500 U/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 (500 U/ml). The Sample Diluent serves as the zero standard (0U/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: Cell culture supernates - Remove particulates by centrifugation and assay immediately or aliquot and store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. 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. 4
3. Duplication of all standards and specimens, although not required, is recommended.
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 SOD 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.

Références pour SOD1 Kit ELISA (ABIN579227)

Karatas, Ozlu, Ozyalvacli, Tosun, Cetinkaya, Donmez, Turker, Bayrakdar: "Intraperitoneal Nigella sativa for prevention of postoperative intra-abdominal adhesions in rats." dans: Journal of investigative surgery : the official journal of the Academy of Surgical Research, Vol. 27, Issue 6, pp. 319-26, (2015) (PubMed).

Lan, Bi, Chen: "Ligustrazine attenuates elevated levels of indoxyl sulfate, kidney injury molecule-1 and clusterin in rats exposed to cadmium." dans: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association, Vol. 63, pp. 62-8, (2014) (PubMed).

Demir, Amanvermez, Kamalı Polat, Karabıçak, Cınar, Kesicioğlu, Polat: "The effect of silymarin on mesenteric ischemia-reperfusion injury." dans: Medical principles and practice : international journal of the Kuwait University, Health Science Centre, Vol. 23, Issue 2, pp. 140-4, (2014) (PubMed).

Détail du antigène

Antigène: SOD1 (Superoxide Dismutase 1, Soluble (SOD1))

Autre désignation: Sod1 (SOD1 Produits)

Synonymes: LOC692639 Kit ELISA, ALS Kit ELISA, ALS1 Kit ELISA, IPOA Kit ELISA, SOD Kit ELISA, hSod1 Kit ELISA, homodimer Kit ELISA, CG11793 Kit ELISA, Cu Kit ELISA, Cu-Zn SOD Kit ELISA, Cu/Zn SOD Kit ELISA, Cu/Zn sod Kit ELISA, Cu/Zn superoxide dismutase Kit ELISA, Cu/ZnSOD Kit ELISA, CuSOD Kit ELISA, CuZn SOD Kit ELISA, CuZn-SOD Kit ELISA, CuZn-SOD1 Kit ELISA, CuZnSOD Kit ELISA, Cu[2+]/Zn[2+]SOD Kit ELISA, Dmel\\CG11793 Kit ELISA, G Kit ELISA, Mn SOD Kit ELISA, SOD-1 Kit ELISA, SOD1 Kit ELISA, Sod-1 Kit ELISA, Sod1 Kit ELISA, To Kit ELISA, To-1 Kit ELISA, Zn SOD Kit ELISA, Zn Sod Kit ELISA, Zn-SOD Kit ELISA, ZnSod Kit ELISA, cSOD Kit ELISA, cSod Kit ELISA, dSOD1 Kit ELISA, l(3)108 Kit ELISA, l(3)68Af' Kit ELISA, l(3)G Kit ELISA, sod Kit ELISA, sod1 Kit ELISA, CU/ZN-SOD Kit ELISA, SODC Kit ELISA, DKFZP469M1833 Kit ELISA, B430204E11Rik Kit ELISA, Cu/Zn-SOD Kit ELISA, Ipo-1 Kit ELISA, Ipo1 Kit ELISA, SOD1L1 Kit ELISA, XSODB Kit ELISA, als Kit ELISA, als1 Kit ELISA, ipoa Kit ELISA, sod1-a Kit ELISA, ZSOD Kit ELISA, cuzn Kit ELISA, Cu/Zn superoxide dismutase Kit ELISA, superoxide dismutase 1 Kit ELISA, Superoxide dismutase 1 Kit ELISA, superoxide dismutase 1, soluble Kit ELISA, superoxide dismutase 1 S homeolog Kit ELISA, Superoxide dismutase [Cu-Zn] Kit ELISA, superoxide dismutase 1 L homeolog Kit ELISA, superoxide dismutase [Cu-Zn]-like Kit ELISA, superoxide dismutase [Cu-Zn] Kit ELISA, superoxide dismutase Sod1 Kit ELISA, SOD Kit ELISA, SOD1 Kit ELISA, Sod1 Kit ELISA, sod1 Kit ELISA, sod1.S Kit ELISA, sod-1 Kit ELISA, sod1.L Kit ELISA, LOC101451855 Kit ELISA, LOC101115136 Kit ELISA

Sujet: Superoxide Dismutases (SODs), originally identified as Indophenoloxidase (IPO), are enzymes that catalyze the converversion of naturally-occuring but harmful superoxide radicals into molecular oxygen and hydrogen peroxide. SOD is a metalloenzyme whose active center is occupied by copper and zinc, sometimes manganese or iron. SOD plays an extremely important role in the protection of all aerobic life-systems, including man, against oxygen toxicity (and the free radicals derived from oxygen). The enzyme superoxide dismutase, or SOD, catalyzes the dismutation of superoxide into oxygen and hydrogen peroxide. SOD is an endogenously produced intracellular enzyme present in essentially every cell in the body. There are at least three forms of superoxide dismutase in nature. Rat erythrocytes contain an SOD enzyme with divalent copper and divalent zinc. Chicken liver mitochondria and E. coli contain a form with trivalent manganese. E. coli also contains a form of the enzyme with trivalent iron. The Cu-Zn enzyme is a dimer of molecular weight 32,500. The two subunits are joined by a disulfide bond. Superoxide dismutases are enzymes that play major roles in the protection of cells against oxidative damage. The two major forms of superoxide dismutase (SOD) in rats are the mitochondrial manganese SOD and the cytosolic copper/zinc SOD. A copper/zinc SOD, isolated from beef liver, has been used intra-articularly for degenerative joint disorders as an anti-inflammatory agent. SOD is also marketed as a nutritional supplement. Cellular SOD is actually represented by a group of metalloenzymes with various prosthetic groups. The prevalent enzyme is cupro-zinc (CuZn) SOD, which is a stable dimeric protein (32,000 D). SOD is an enzyme associated with copper, zinc, and manganese by body cells, and breaks down the superoxide free radicals. It is said that SOD protects the lens of the eyes by guarding against free radical damage.

ID gène: 3110

Pathways: Sensory Perception of Sound, Transition Metal Ion Homeostasis