Donkey Anti-Guinea Pig IgG/PE-Cy7

The Donkey Anti-Guinea Pig IgG/PE-Cy7 antibody is a versatile reagent for various applications, including Immunofluorescence (IF) and Flow Cytometry (FCM). Below are the protocols for each validated application.

For IF, the following steps are recommended:

- Reagents or Buffers: Phosphate-buffered saline (PBS), 4% paraformaldehyde for fixation, and a mounting medium with DAPI for nuclear staining.

- Sample Preparation: Fix cells or tissue sections with 4% paraformaldehyde, permeabilize if necessary, and block with a suitable blocking buffer.

- Antibody Incubation: Dilute the Donkey Anti-Guinea Pig IgG/PE-Cy7 antibody to 1:100-1000 in the blocking buffer and incubate for 1-2 hours at room temperature or overnight at 4°C.

- Washing or Detection: Wash samples thoroughly with PBS, and proceed with mounting using a DAPI-containing mounting medium.

- Technical Notes: Optimize the antibody dilution and incubation time based on the specific experimental conditions and the intensity of the fluorescence signal.

For FCM, follow these guidelines:

- Reagents or Buffers: PBS, FACS buffer (PBS with 1% BSA and 0.1% sodium azide), and a fixation buffer if necessary.

- Sample Preparation: Prepare single-cell suspensions, and if necessary, fix and permeabilize the cells according to the experimental requirements.

- Antibody Incubation: Dilute the Donkey Anti-Guinea Pig IgG/PE-Cy7 antibody to 1:100-1000 in FACS buffer and incubate for 30 minutes to 1 hour on ice.

- Washing or Detection: Wash cells with FACS buffer, and proceed with flow cytometric analysis.

- Technical Notes: Ensure proper compensation for the PE-Cy7 channel, and consider using isotype controls to verify specificity.

Use assay conditions validated for IF, apply Donkey Anti-Guinea Pig IgG/PE-Cy7 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-Guinea Pig IgG/PE-Cy7 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.