| Field | Specification |
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Product Overview
The 96-Well Super High Binding ELISA Plates are designed to give strong ELISA signals. Strong protein binding to the plate surface increases ELISA sensitivity.
In the indirect format, the antigen is coated directly onto the plate, the plate is blocked, an antibody against the antigen is added, and the bound antibody is detected with an enzyme-linked secondary antibody. When two antibodies (A and B) against the antigen are available, antibody A can be coated first to capture the antigen, which is then detected with antibody B and an enzyme-linked antibody; this antibody pre-coating may further increase sensitivity.
Each pack contains 4 plates. Black plates give a low background for fluorescence detection. Transparent plates are for detection by light absorbance.
Product Description
| Option | Black (fluorescence detection) | Transparent (absorbance detection) |
|---|---|---|
| Catalog # | ELISA96B | ELISA96T |
| Plate Color | Black | Transparent |
| Detection | Fluorescence | Light absorbance |
| Contents | 4 plates | 4 plates |
Specifications
| Format | 96-well plates |
|---|---|
| Plate Color | Black or transparent (two options; see Product Description) |
| Pack Size | 4 plates per pack |
| Typical Coating Volume | 100 µl per well |
| Typical Wash / Block Volume | 360 µl per well |
| Catalog # | ELISA96B, ELISA96T |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped at room temperature. |
Protocol Outline
- Coat each well with 100 µl of antigen (2-fold dilution series in PBS with 0.02% sodium azide; a typical top concentration is 1 µg/ml) and rotate at 4°C overnight.
- Wash 4 times with 360 µl of PBS with 0.01% Tween-20, then block with 360 µl of PBS with 0.02% sodium azide and 3% fetal bovine serum for 2 hours at room temperature.
- Wash, add 100 µl of primary antibody diluted in PBS with 0.02% sodium azide and 0.1% fetal bovine serum, and incubate with rotation for 1 hour at room temperature.
- Wash, add 100 µl of the secondary antibody for 1 hour, wash again, add 200 µl of detection solution and read absorbance or fluorescence.
The full ProFoldin protocol “96-well Super High Binding ELISA Plates” is linked under Documents.
Notes
- For antibody pre-coating, coat 100 µl of 1 µg/ml antibody A at 4°C overnight or at 37°C for 2 hours, block, and then continue from the antigen step without a second blocking step.
- In the antibody pre-coating format, antibody B should come from a different host species than antibody A, or be directly labeled, so that the enzyme-linked secondary antibody does not bind the capture antibody.
- Sodium azide inhibits horseradish peroxidase (HRP); leave it out of buffers used with HRP-conjugated antibodies.
Safety & Handling
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