Rat Estrogen ELISA Kit

Standard Curve Range: 3ng/L-190ng/L

Sensitivity: 1.92ng/L

Size: 96 wells

Storage: Store the reagents at 2-8°C. For long term storage refer to the expiration date keep it at -20°C. Avoid repeated thaw cycles.

*This product is for research use only, not for use in diagnosis procedures. It’s highly recommend to read this instruction entirely before the use.

PRECICION

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

CV(%) = SD/mean x 100

Intra-Assay: CV<8%

Inter-Assay: CV<10%

INTENDED USE

This sandwich kit is for the accurate quantification of Rat Estrogen(also known as E)in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

ASSAY PRINCIPLE

This kit is a Enzyme-Linked Immunosorbent Assay (ELISA). E is added to the wells pre-coated with E monoclonal antibody. After cubation a biotin-conjugated anti-Rat E antibody is added and binds to Rat E. After incubation unbound biotin-conjugated anti-Rat E antibody is washed away during a washing step. Streptavidin-HRP is added and binds to the biotin-conjugated anti-Rat E antibody. After incubation unbound Streptavidin-HRP is washed away during a washing step. Substrate solution is then added and color develops in proportion to the amount of Rat E. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.

MATERIALS REQUIRED BUT NOT SUPPLIED

1.37°C±0.5°C incubator

2.Microplate reader with 450 ± 10nm wavelength filter

3.Precision pipettes and disposable pipette tips

4.Clean tubes

5.Deionized or distilled water

6.Absorbent paper

PRECAUTIONS

1.Prior to use, the kit and sample should be warmed naturally to room temperature 30 minutes.

2.Once the desired number of strips has been removed, immediately reseal the bag to protect the remain from deterioration. Cover all reagents when not in use.

3.Make sure pipetting order and rate of addition from well-to-well when pipetting reagents.

4.This instruction must be strictly followed in the experiment.

5.Pipette tips and plate sealer in hand should be clean and disposable to avoid cross-contamination.

6.Avoid using the reagents from different batches together.

7.Substrate solution B is sensitive to light, don’t expose substrate solution B to light for a long time.

8.Stop solution contains acid. Please wear eye, hand and skin protection when using this material. Avoid contact of skin or mucous membranes with kit reagent.

SPECIMEN COLLECTION

Serum: Allow serum to clot for 10-20 minutes at room temperature. Centrifuge at 2000-3000 RPM for 20 minutes.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C within 30 minutes of collection.

Urine: Collect by sterile tube. Centrifuge at 2000-3000 RPM for approximately 20 minutes. When collecting pleuroperitoneal fluid and cerebrospinal fluid, please follow the procedures above-mentioned.

Cell culture supernatant: Collect by sterile tubes when examining secrete components. Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect the supernatants carefully. When examining the components within the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the cell concentration of approximately 1 million/ml. Damage cells through repeated freeze-thaw cycles to let out the inside components. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

Tissue: Rinse tissues in PBS(pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes.

Rebecca Yan

Product Manager
Span Biotech Ltd.
Tel: +86(755)89589611
Cell Phone:+8618823462100(WhatsApp)
Web:www.spanbio.com