Acid Phosphatase Assay Kit User Manual Catalog
Contents
1. please go online Vill NOTES FOR RESEARCH USE ONLY NOT FOR S OR THERAPEUTIC PROCEDURES2. Acid Phosphatase Assay Kit User Manual Catalog MBS822357 Detection and Quantification of Acid Phosphatase 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 k INTRODU CHON em 2 ll 5 aa E e SSe 3 11 MATERIALS REQUIRED BUT NOT PROVIDED eese emen 3 SAMPLE PREPARATION enar hae oremus pera ko ru negar keen est Redes Vn 4 M ASSAY PROCEDURE rire i e tenera tame 5 CALCULATION E 6 VII TECHNICAL doe rentre db ru eh e eite triente 7 VIN eji MM A 2050521505 7 FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES l INTRODUCTION Acid phosphatases AP dephosphorylate phosphate groups from phosphate esters under acid conditions Different acid phosphatase isozymes are found in different organs and their serum levels are used as a diagnostic for disease in the corresponding organs Elevated prostatic acid phosphatase levels may indicate the presence of prostate cancer and elevated tartrate resistant acid phosphatase levels may indicate bone disease FOR RESEA
3. RCH USE ONLY NOT FOR USE I NOSTIC OR THERAPEUTIC PROCEDURES KIT COMPONENTS Component 96 Well Microplate Reagent I Reagent II Reagent III Reagent IV Standard Technical Manual Note Volume 30ml x2 5mlx1 10mlx1 15 mix 1 1mlx1 1 mg ml 1 Manual ON Standard add 9 ml distilled water to dilute as the Standard Solution before use the concentration is 0 1 mg ml Ill MATERIALS REQUIRED NOT PROVIDED 1 Microplate reader to read absorbance at 510 nm 2 Distilled water 3 Pipettor 4 Pipette tips 5 Mortar 6 Ice 7 Centrifuge for cell lysis 8 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 0 5ml Reagent 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 5ml Reagent 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 FOR RESEARCH USE ONLY NOT FOR USE I NOSTIC OR THERAPEUTIC PROCEDURES V ASSAY PROCEDURE Add following reagents in
4. the microplate Reagent Experiment Standard Blank Control Sample 10 ul Standard Solution 10 ul Distilled water 10 ul Reagent 40 ul 40 ul 40 ul 40 ul Mix put it in water bath of 37 C for 5 minutes Reagent II 40 ul 40 ul 40 ul 40 ul Mix put it in water bath of 37 C for 60 minutes Reagent III 80 ul 80 ul 80 ul 80 ul Reagent IV 120 ul 120 ul 120 ul 120 ul Sample 10 ul Mix record absorbance measured at 510 nm Notice 1 Keep Reagent 111 Reagent IV in dark 2 When Reagent IV turn to blue it is can not use 3 After add Reagent IV mix immediately 4 The sample should be prepared freshly For serum sample add 10 mg Disodium Citrate or 5 mg Sodium Acid Sulfate in 1 ml serum store at 4 FOR RESEARCH USE ONLY NOT FOR OR THERAPEUTIC PROCEDURES VI CALCULATION 1 Calculation of AP in cell tissue sample AP U mg prot 0 1 x ODsample 7 OD controt OD standard OD Cpr protein concentration mg mL The protein concentration should be detected separately you can buy BCA protein concentration assay kit 2 Calculation of AP in serum sample AP U 100ml 0 1 ODsample D OD OD standard 100 FOR RESEARCH USE ONLY FOR OR THERAPEUTIC PROCEDURES Vil TECHNICAL SUPPORT For troubleshooting information or assistance
Download Pdf Manuals
Related Search