- Product Name
- Rat Prl(Prolactin) ELISA Kit
- Alias
- Prolactin ELISA Kit, PRL ELISA Kit, PRL ELISA Kit
- Catalogue No.
- ER0076
- Size
- 48T/96T
- Species
- Rat
- UniProt No.
- P01237
- Sample Type
- Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
- Detection Method
- Sandwich ELISA, Double Antibody
- Detection Wavelength
- OD450
- Reaction Duration
- 4 hours
- Range
- 0.781-50ng/ml
- Sensitivity
- 0.469ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with PRL , no obvious cross reaction with other analogues.
- ELISA Kit Components
| Kit Components | Item | Size(48T) | Size(96T) | Storage Condition for Opened Kit |
| E001 | ELISA Microplate(Dismountable) | 8×6 | 8×12 | Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C |
| E002 | Lyophilized Standard | 1vial | 2vial | Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C |
| E003 | Biotin-labeled Antibody(Concentrated, 100X) | 60ul | 120ul | 2-8°C (Avoid Direct Light) |
| E034 | HRP-Streptavidin Conjugate(SABC, 100X) | 60ul | 120ul |
| E024 | TMB Substrate | 5ml | 10ml |
| E039 | Sample Dilution Buffer | 10ml | 20ml | 2-8°C |
| E040 | Antibody Dilution Buffer | 5ml | 10ml |
| E049 | SABC Dilution Buffer | 5ml | 10ml |
| E026 | Stop Solution | 5ml | 10ml |
| E038 | Wash Buffer(Concentrated, 25X) | 15ml | 30ml |
| E006 | Plate Sealer | 3 pieces | 5 pieces | |
| E007 | Product Description | 1 copy | 1 copy |
- Required Instruments and Reagents
-
- Microplate reader (wavelength: 450nm)
- 37°C incubator (CO2 incubator for cell culture is not recommenced.)
- Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
- Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
- Sterile tubes and Eppendorf tubes with disposable tips
- Absorbent paper and loading slot
- Deionized or distilled water
- Assay Procedure Summary
-
- Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
- Washing: Wash the plate twice without immersing.
- Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
- Washing: Wash the plate three times and immerse for 1min each time.
- Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
- Washing: Wash the plate five times and immerse for 1min each time.
- Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
- Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
- Standard Curve
-
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)
Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
| STD.(ng/ml) | OD-1 | OD-2 | Average |
| 0 | 0.111 | 0.115 | 0.113 |
| 0.781 | 0.241 | 0.249 | 0.245 |
| 1.562 | 0.39 | 0.402 | 0.396 |
| 3.125 | 0.55 | 0.566 | 0.558 |
| 6.25 | 0.795 | 0.819 | 0.807 |
| 12.5 | 1.036 | 1.066 | 1.051 |
| 25 | 1.666 | 1.714 | 1.69 |
| 50 | 2.372 | 2.44 | 2.406 |
- Recovery
-
Add a certain amount of PRL into the sample. Calculate the recovery by comparing the measured value with the expected amount of PRL in the sample.
| Sample Type | Recovery Range(%) | Average(%) |
| serum(n=10) | 88-104 | 97 |
| EDTA plasma(n=10) | 88-103 | 96 |
| Heparin plasma(n=10) | 86-95 | 92 |
- Linearity
-
Dilute the sample with a certain amount of PRL at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 85-103% | 91-105% | 89-104% |
| EDTA plasma(n=10) | 88-98% | 87-101% | 89-101% |
| Heparin plasma(n=10) | 81-99% | 82-98% | 80-98% |
- Precision(%)
-
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
| Item | Intra-assay Precision | Inter-assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/ml) | 1.57 | 5.83 | 24.13 | 1.48 | 6.04 | 24.69 |
| Standard deviation | 0.09 | 0.25 | 1.32 | 0.06 | 0.32 | 1.2 |
| CV(%) | 5.5 | 4.34 | 5.46 | 4.25 | 5.37 | 4.88 |
- Stability
-
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
| Average(%) | 80 | 95-100 |
Manual