GOT2 ELISA Kit

Catalog No. ABIN6974258

Product Details GOT2 ELISA Kit

Target: GOT2 (Glutamic-Oxaloacetic Transaminase 2, Mitochondrial (Aspartate Aminotransferase 2) (GOT2))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 1.56 mIU/mL - 100 mIU/mL

Minimum Detection Limit: 1.56 mIU/mL

Application: ELISA

Purpose: For the quantitative determination of mouse aspartate aminotransferase (AST) concentrations in serum, plasma, cell culture supernates.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of mouse AST. No significant cross-reactivity or interference between mouse AST and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between mouse AST and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 0.39 mIU/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 100 mIU/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (100 mIU/mL). Sample Diluent serves as the zero standard (0 mIU/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for GOT2 ELISA Kit (ABIN6974258)

Xiahou, Wang, Shen, Zhu, Xu, Hu, Guo, Li, Tian, Liu, Liang: "NMI and IFP35 serve as proinflammatory DAMPs during cellular infection and injury." in: Nature communications, Vol. 8, Issue 1, pp. 950, (2018) (PubMed).

Song, Kong, Zhang, Zhou, Li: "Ulinastatin Protects against CVB3-Induced Acute Viral Myocarditis through Nrf2 Activation." in: Inflammation, Vol. 41, Issue 3, pp. 803-810, (2018) (PubMed).

Chen, Zhou, Wu, Wang, Wang: "FTO-dependent function of N6-methyladenosine is involved in the hepatoprotective effects of betaine on adolescent mice." in: Journal of physiology and biochemistry, (2015) (PubMed).

Sánchez-Cordón, Pérez de Diego, Gómez-Villamandos, Sánchez-Vizcaíno, Pleguezuelos, Garfia, del Carmen, Pedrera: "Comparative analysis of cellular immune responses and cytokine levels in sheep experimentally infected with bluetongue virus serotype 1 and 8." in: Veterinary microbiology, Vol. 177, Issue 1-2, pp. 95-105, (2015) (PubMed).

Target Details for GOT2

Target: GOT2 (Glutamic-Oxaloacetic Transaminase 2, Mitochondrial (Aspartate Aminotransferase 2) (GOT2))

Alternative Name: glutamic-oxaloacetic transaminase 2, mitochondrial (aspartate aminotransferase 2) (GOT2 Products)

Synonyms: KAT4 ELISA Kit, KATIV ELISA Kit, mitAAT ELISA Kit, AI789014 ELISA Kit, Got-1 ELISA Kit, cAspAT ELISA Kit, cCAT ELISA Kit, AL022787 ELISA Kit, FABP-pm ELISA Kit, Got-2 ELISA Kit, mAspAT ELISA Kit, ASPATA ELISA Kit, got2 ELISA Kit, zgc:66329 ELISA Kit, fj40h07 ELISA Kit, wu:fj40h07 ELISA Kit, zgc:56425 ELISA Kit, glutamic-oxaloacetic transaminase 2 ELISA Kit, glutamic-oxaloacetic transaminase 1, soluble ELISA Kit, glutamatic-oxaloacetic transaminase 2, mitochondrial ELISA Kit, glutamic-oxaloacetic transaminase 2a, mitochondrial ELISA Kit, glutamic-oxaloacetic transaminase 2 L homeolog ELISA Kit, Aspartate amino transferase activity ELISA Kit, glutamic-oxaloacetic transaminase 2b, mitochondrial ELISA Kit, GOT2 ELISA Kit, got2 ELISA Kit, Got1 ELISA Kit, Got2 ELISA Kit, got2a ELISA Kit, got2.L ELISA Kit, AST ELISA Kit, got2b ELISA Kit

Background:

Abbreviation: GOT2

Alias: FLJ40994, KAT4, KATIV, mitAAT, aspartate aminotransferase 2|kynurenine aminotransferase IV

UniProt: P05202

Pathways: Monocarboxylic Acid Catabolic Process