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Click Here to Download This Entire GuideChoose the detection enzyme  |
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| Choose the appropriate ABC kit |
To detect a biotinylated target, you will need:
For example, to detect a primary antibody made in rabbit, the appropriate choice is a VECTASTAIN® ABC Kit designated Rabbit IgG. |
| Choose a convenient format | VECTASTAIN® ABC Kits are available in economical concentrated formats. For additional convenience and ease of use some peroxidase-based kits are offered in ready-to-use prediluted stabilized formats. |
Tumor: • Ki67 (m), VECTASTAIN® Elite® ABC Kit, Vector® DAB (brown) • CD34 (m), VECTASTAIN® ABC-AP Kit, Vector® Blue (blue) • Cytokeratin AE1/AE3 (m), VECTASTAIN® ABC-AP Kit, Vector® Red (red).
Consider Species Cross-Reactivity
When choosing the optimal detection system for your application, it is important to consider not only the species of the primary antibody but also the species of the tissue under examination. If the species of the primary antibody and the species of the tissue are closely related (for example, rat and mouse), the biotinylated secondary antibody may cross-react with endogenous IgG in the tissue section. This can lead to background staining.
The following options may minimize background staining in these instances:
Substrates
After choosing the VECTASTAIN® ABC Kit for your application, select a substrate that matches the enzyme system of the kit (p. 26-29).
Endometrium: • Progesterone Receptor (rm), VECTASTAIN® Universal ABC- AP Kit, Vector® Blue AP Substrate (blue) • CD34 (m), VECTASTAIN® Universal Elite® ABC Kit, Vector® DAB HRP Substrate (brown).
Tumor: • p53 protein (m), VECTASTAIN® ABC-AP Kit, Vector® Red AP Substrate (red) • Pan-Cytokeratin (sheep), VECTASTAIN® Elite® ABC Kit, Vector® SG HRP Substrate (blue/gray).
Intestine: • Desmin (m), VECTASTAIN® Elite® ABC Kit, ImmPACT™ DAB (brown) substrate. • Cytokeratin (m), VECTASTAIN® Elite® ABC Kit, Vector® VIP (purple) substrate.
Tonsil: • Cyclin A (m), VECTASTAIN® Universal ABC-AP Kit, Vector® Blue AP Substrate (blue) • CD20 (m), VECTASTAIN® Universal Elite® ABC Kit, Vector® NovaRED™ HRP Substrate (red).
Breast Carcinoma: • Estrogen Receptor (m); VECTASTAIN® Elite® ABC Kit, Vector® NovaRED substrate (red) • CD34 (m), VECTASTAIN® Elite® ABC Kit, DAB substrate (brown) • Cytokeratin 8/18 (m), VECTASTAIN® Elite® ABC Kit, Vector® SG substrate (blue/gray).
Tonsil: • CD3 (m), VECTASTAIN® Universal Elite® ABC Kit, Vector® VIP substrate (purple) • CD20 (m), VECTASTAIN® Universal Elite® ABC Kit, Vector® SG substrate (blue/gray).
