Aromatase ELISA Kit

Catalog No. ABIN6975844

Product Details Aromatase ELISA Kit

Target: Aromatase (CYP19A1) (Cytochrome P450, Family 19, Subfamily A, Polypeptide 1 (CYP19A1))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 6.25 pg/mL - 400 pg/mL

Minimum Detection Limit: 6.25 pg/mL

Application: ELISA

Purpose: For the quantitative determination of mouse aromatase (ARO) concentrations in serum, plasma, tissue homogenates.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

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

Sensitivity: 1.56 pg/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 400 pg/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 (400 pg/mL). Sample Diluent serves as the zero standard (0 pg/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).

Note:

Serum and plasma samples require a 4-fold dilution into Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.

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

Target Details for Aromatase

Target: Aromatase (CYP19A1) (Cytochrome P450, Family 19, Subfamily A, Polypeptide 1 (CYP19A1))

Alternative Name: cytochrome P450, family 19, subfamily A, polypeptide 1 (CYP19A1 Products)

Synonyms: ARO ELISA Kit, ARO1 ELISA Kit, CPV1 ELISA Kit, CYAR ELISA Kit, CYP19 ELISA Kit, CYPXIX ELISA Kit, P-450AROM ELISA Kit, Aromatase ELISA Kit, Cyp19 ELISA Kit, Cyp19a ELISA Kit, p450arom ELISA Kit, P450 arom ELISA Kit, aro ELISA Kit, cyp19 ELISA Kit, cyp19a ELISA Kit, p450arom-A ELISA Kit, P450AROM ELISA Kit, Ar ELISA Kit, ArKO ELISA Kit, Int-5 ELISA Kit, Int5 ELISA Kit, CYP19P1 ELISA Kit, CYP19A1 ELISA Kit, CYP19a1 ELISA Kit, P450aromA ELISA Kit, ar1 ELISA Kit, cypXIX ELISA Kit, etID309862.18 ELISA Kit, zgc:136541 ELISA Kit, AroB ELISA Kit, CYP19a2 ELISA Kit, P450aromB ELISA Kit, cyp19b ELISA Kit, zgc:92614 ELISA Kit, cyp19a1 ELISA Kit, cytochrome P450 family 19 subfamily A member 1 ELISA Kit, cytochrome P450, family 19, subfamily a, polypeptide 1 ELISA Kit, cytochrome P450 family 19 subfamily A member 1 L homeolog ELISA Kit, aromatase ELISA Kit, cytochrome P450 aromatase ELISA Kit, cytochrome P450, family 19, subfamily A, polypeptide 1 ELISA Kit, cytochrome P450 19A1 ELISA Kit, cytochrome P450, family 17, subfamily A, polypeptide 1 ELISA Kit, cytochrome P450, family 19 ELISA Kit, cytochrome P450, family 19, subfamily A, polypeptide 1a ELISA Kit, cytochrome P450, family 19, subfamily A, polypeptide 1b ELISA Kit, cytochrome P450 19A1-like ELISA Kit, CYP19A1 ELISA Kit, Cyp19a1 ELISA Kit, cyp19a1.L ELISA Kit, SAV_2383 ELISA Kit, pfcyp19b-tstr ELISA Kit, CYP19 ELISA Kit, cyp19a1a ELISA Kit, cyp19a1b ELISA Kit, LOC101174651 ELISA Kit

Background:

Abbreviation: CYP19A1

Alias: ARO, ARO1, CPV1, CYAR, CYP19, MGC104309, P-450AROM, OTTHUMP00000162543|aromatase|cytochrome P450, subfamily XIX (aromatization of androgens)|estrogen synthetase|flavoprotein-linked monooxygenase|mic

UniProt: P28649

Pathways: Metabolism of Steroid Hormones and Vitamin D, Steroid Hormone Biosynthesis, C21-Steroid Hormone Metabolic Process, Carbohydrate Homeostasis