Donkey Anti-Rabbit IgG/APC

The Donkey Anti-Rabbit IgG/APC is a versatile antibody conjugate suitable for various applications, including Immunofluorescence (IF) and Flow Cytometry (FCM).

For IF, start by preparing your samples, either by fixing cells with a suitable fixative such as paraformaldehyde or by using frozen tissue sections.

Reagents or buffers needed include the antibody conjugate, a blocking buffer to reduce non-specific binding, and a mounting medium if necessary.

Sample preparation involves permeabilization if intracellular targets are of interest, followed by blocking to prevent non-specific binding.

Antibody incubation typically involves diluting the Donkey Anti-Rabbit IgG/APC 1:100 to 1:1000 in an appropriate buffer and incubating for a period that allows sufficient binding, usually 1 hour at room temperature or overnight at 4°C.

Washing steps are crucial to remove unbound antibody, using a buffer such as PBS.

Detection in IF involves observing the fluorescence signal under a fluorescence microscope.

Technical notes: Optimize the dilution of the primary antibody and the conjugate for best results, and consider using controls to validate specificity.

For FCM, prepare your cell samples by washing and resuspending them in an appropriate buffer.

Reagents needed include the Donkey Anti-Rabbit IgG/APC, FACS buffer, and potentially a fixation step post-staining.

Sample preparation involves staining cells with a primary rabbit antibody against the target of interest, followed by incubation with the Donkey Anti-Rabbit IgG/APC at a dilution of 1:100 to 1:1000 for 30 minutes to 1 hour on ice.

Washing steps are essential to remove unbound conjugate, using a buffer like PBS.

Detection in FCM involves analyzing the stained cells using a flow cytometer.

Technical notes: Use appropriate controls, such as isotype controls for the primary antibody and a secondary antibody-only control, to validate the specificity of staining.

Always optimize the concentration of the conjugate and the primary antibody for the specific application and cell type being studied.

Use assay conditions validated for IF, apply Donkey Anti-Rabbit IgG/APC at the recommended working dilution, include proper positive and negative controls, perform thorough wash steps, and document incubation time, temperature, and detection conditions for reproducible performance. Recommended dilution: FCM=1:100-1000;

IF=1:100-1000.

Use assay conditions validated for FCM, apply Donkey Anti-Rabbit IgG/APC at the recommended working dilution, include proper positive and negative controls, perform thorough wash steps, and document incubation time, temperature, and detection conditions for reproducible performance. Recommended dilution: FCM=1:100-1000;

IF=1:100-1000.