Donkey Anti-Rabbit IgG/AP

The Donkey Anti-Rabbit IgG/AP antibody is a versatile reagent that can be used in various applications, including Western Blotting (WB), Enzyme-Linked Immunosorbent Assay (ELISA), Immunohistochemistry on Paraffin-embedded tissues (IHC-P), and Immunohistochemistry on Frozen tissues (IHC-F). The following protocols are recommended for each application.

Reagents or buffers: Tris-buffered saline with Tween 20 (TBST), skim milk or bovine serum albumin (BSA) for blocking, and a suitable substrate for alkaline phosphatase (AP) detection.

Sample preparation: Separate proteins by SDS-PAGE and transfer them to a membrane.

Antibody incubation: Incubate the membrane with the Donkey Anti-Rabbit IgG/AP antibody at a dilution of 1:1000-10000 in TBST with 5% skim milk or 5% BSA for 1-2 hours at room temperature or overnight at 4°C.

Washing or detection: Wash the membrane three times with TBST, then detect the signal using a suitable AP substrate.

Technical notes: Optimize the blocking conditions and antibody dilution to minimize background and maximize specific signal.

Reagents or buffers: Coating buffer (e.g., carbonate-bicarbonate buffer), blocking buffer (e.g., PBS with 1% BSA), and a suitable substrate for AP detection.

Sample preparation: Coat the ELISA plate with the antigen of interest, then block non-specific binding sites.

Antibody incubation: Incubate the plate with the Donkey Anti-Rabbit IgG/AP antibody at a dilution of 1:1000-10000 in blocking buffer for 1-2 hours at room temperature.

Washing or detection: Wash the plate three times with PBS-T, then detect the signal using a suitable AP substrate.

Technical notes: Optimize the coating and blocking conditions to minimize non-specific binding.

Reagents or buffers: Xylene, ethanol, antigen retrieval buffer (if necessary), and a suitable substrate for AP detection.

Sample preparation: De-paraffinize and rehydrate tissue sections, then perform antigen retrieval if necessary.

Antibody incubation: Incubate the sections with the Donkey Anti-Rabbit IgG/AP antibody at a dilution of 1:100-1000 in a suitable buffer (e.g., PBS with 1% BSA) for 1-2 hours at room temperature or overnight at 4°C.

Washing or detection: Wash the sections three times with PBS, then detect the signal using a suitable AP substrate.

Technical notes: Optimize the antigen retrieval and antibody dilution to minimize background and maximize specific signal.

Reagents or buffers: Acetone or methanol for fixation, and a suitable substrate for AP detection.

Sample preparation: Fix tissue sections with acetone or methanol, then permeabilize if necessary.

Antibody incubation: Incubate the sections with the Donkey Anti-Rabbit IgG/AP antibody at a dilution of 1:100-1000 in a suitable buffer (e.g., PBS with 1% BSA) for 1-2 hours at room temperature or overnight at 4°C.

Washing or detection: Wash the sections three times with PBS, then detect the signal using a suitable AP substrate.

Technical notes: Optimize the fixation and antibody dilution to minimize background and maximize specific signal.