The ImmPRESS polymerized reporter enzyme staining system uses novel conjugation and micropolymer chemistries to create a highly sensitive, ready-to-use, one-step, biotin-free detection system. This unique micropolymer of highly active enzyme (alkaline phosphatase) is attached to our affinity purified secondary antibodies, producing reagents with outstanding sensitivity and low background.
The ImmPRESS (15 ml) Kit will stain approximately 75-150 sections.
| Unit Size | 15 ml |
|---|---|
| Applications | Immunohistochemistry / Immunocytochemistry, In situ hybridization, ELISAs |
| Technology | Micropolymer Reagents |
| Detection Enzyme(s) | Alkaline Phosphatase |
| Conjugate | Micropolymer AP |
| Target Species | Rat |
| Format | Ready-to-Use |
The ImmPRESS Excel kits are a two-step peroxidase polymer based detection system that include an unconjugated amplifier antibody. This intermediate amplifier antibody increases sensitivity of the assay at least three- to four-fold over that of a one-step polymer based system, by facilitating the introduction of more peroxidase enzyme at the site of specific antigen localization. This increase in sensitivity would be advantageous in instances of weak antigen expression, and to further dilute out an expensive primary antibody. The ImmPRESS Excel Amplifier kits are presented as a complete kit format that include enzyme quench, blocking serum and ImmPACT® DAB EqV peroxidase substrate, in addition to the amplifier antibody and defined ImmPRESS polymer secondary antibody. However, as the two published references below indicate, this format is still modular and allows for the substitution of different detection reagents. These references describe using ImmPACT NovaRED™ peroxidase substrate in combination with the ImmPRESS Excel Amplifier kits. Oncogenesis (2017) 6, e293; doi:10.1038/oncsis.2016.82 Tan, E.M.S., et al (2017) Front. Med. 4:162 (doi: 10.3389/fmed.2017.00162)
Yes, we conducted studies on the compatibility of the ImmPRESS HRP polymer reagents with three commercially available autostainers (Agilent/Dako Autostainer Plus, Leica Bond Rx, and Ventana Discovery Ultra). We showed that our reagents are suitable for IHC detection on each of these platforms. The ImmPRESS polymer reagents and enzyme substrates generated equivalent IHC staining results compared to reagents from the instrument manufacturer. Some modifications in the protocol were performed to optimize the signal-to-noise ratio, such as a shorter incubation time for the ImmPRESS polymer and increase in the number of buffer washes following polymer incubation. Please see the following application note for more details about each automated platform:https://vectorlabs.com/wp-content/uploads/2022/11/VL_LIT3023_ImmPRESS_AppNote_4.pdf
The staining procedure is simple as shown in the diagram below. Following a blocking step with the diluted normal horse serum, sections are incubated with primary antibody. After a brief wash, the appropriate ImmPRESS Reagent is added to the sections and incubated for 30 minutes. Sections are again rinsed and the slides are developed with the peroxidase substrate of choice.
Applicable patents and legal notices are available at legal notices.
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