Hydrogen Peroxide Assay Kit User Manual Catalog
Contents
1. 0 062 Vst 8 06 x A1 A2 0 08 7 W W fresh weight of sample g Vst total volume of extract 0 5ml 3 According to concentration of cell or bacteria umol 10 AT A2 0 08 0 062 200 Vst 0 04 x A1 A2 0 08 200 Quantity of cell or bacteria 200x10 FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES Vil TECHNICAL SUPPORT For troubleshooting information or assistance please go online Vill NOTES FOR RESEARCH USE ONLY NOT FOR S OR THERAPEUTIC PROCEDURES2. FOR RESEARCH USE ONLY NOT FOR USE I NOSTIC OR THERAPEUTIC PROCEDURES KIT COMPONENTS Component 96 Well Microplate Reagent I Reagent II Reagent III Technical Manual Note Volume Powder x 1 15 ml x 2 30 ml x2 1 Manual Reagent I add 10ml hydrochloric acid to dissolve it absolutely before use stored at 4 C Ill MATERIALS REQUIRED BUT NOT PROVIDED 1 Microplate reader to read absorbance at 415 nm 2 Acetone 60ml 3 Hydrochloric acid 10ml 4 Pipettor 5 Pipette tips 6 Mortar 7 Ice 8 Centrifuge for cell lysis 9 Timer FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES IV SAMPLE PREPARATION 1 For cell and bacteria samples Collect cell or bacteria into centrifuge tube discard the supernatant after centrifugation add 500 ul acetone for 2x10 cell or bacteria sonicate with power 2096 sonication 3s intervation 10s repeat 30 times centrifuged at 8000g 4 C for 10min take the supernatant into a new centrifuge tube and keep it on ice for detection 2 For tissue samples Weight 0 05g tissue homogenize with 0 5 ml acetone on ice centrifuged at 8000g 4 C for 10min take the supernatant into a new centrifuge tube and keep it on ice for detection 3 For serum or plasma samples Detect directly Note 1 Acetone should be preco led before use because of its volatile grind the sample on ice 2 Please wear disposable gloves and masks because of the high vo
3. Hydrogen Peroxide Assay Kit User Manual Catalog MBS822356 Detection and Quantification of Hydrogen Peroxide Concentrations in Urine Serum Plasma Other biological fluids Tissue Extracts Cell Lysate Cell culture media Samples For research use only Not for diagnostic or therapeutic procedures FOR RESEARCH USE ONLY NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES LOINTRODUCTION s 2 Ihe KIT COMPONENTS 3 Ill MATERIALS REQUIRED BUT NOT PROVIDED esee 3 Vi SAMPLE PREPARATION 2 t ee Du epa 4 M ASSAY PROCEDURE ecce aeo nre e erreur petere ned eap eei anna reine nu ue 5 VI CALCULATION Mt 6 TECHNICAL SUPPONRLT eati tiet tot rettet ea canted ii inrer netta nerve 7 NOTES ro tritt ee tta oit ences teen eren prr al i Nene 7 FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES l INTRODUCTION Hydrogen Peroxide H202 is a reactive oxygen metabolic byproduct that serves as key regulator for a number of oxidative stress related states Functioning through NF kappaB and other factors hydroperoxide mediated pathways have been linked to asthma inflammatory arthritis atherosclerosis diabetic vasculopathy osteoporosis neuro degenerative diseases Down s syndrome and immune system diseases
4. latile of reagents in the kit FOR RESEARCH USE ONLY NOT FOR USE I NOSTIC OR THERAPEUTIC PROCEDURES V ASSAY PROCEDURE Preheat Reagent I Il and IIl on 37 C only for mammal or 25 C for general organisms for 10 minutes Reagent Experiment Control Sample all the supernatant Acetone 500 ul Reagent 50 ul 50 ul Reagent II 100 ul 100 ul Centrifuged at 4000 g 25 C for 10 minutes discard the supernatant leave the precipitation Reagent III 500 ul 500 ul Mix stand at room temperature for 5 minutes take 200 ul into the microplate record absorbance measured at 415 nm absorbances of experiment and control are denoted as 1 and A2 do th control only once FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES VI CALCULATION 1 The regression equation under standard condition Y 0 062X 0 08 X concentration of standard Y absolute absorbance A1 A2 2 Calculation of H202 in serum or plasma umol mL A1 2 0 08 0 062 16 13 x A1 2 0 08 3 Calculation of H2072 in cell bacteria and tissue 1 According to protein concentration of sample H20 umol mg prot A1 A2 0 08 0 062 Cpr Cpr protein concentration mg ml The protein concentration should be detected separately you can buy BCA protein concentration assay kit 2 According to fresh weight of sample H202 umol g FW A1 A2 0 08
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