Overview of WDR91 and Antibody Specifications

WDR91 (WD Repeat Domain 91) is a protein characterized by multiple WD40 repeat motifs, which typically serve as scaffolding platforms mediating protein–protein interactions. WDR91 has been implicated in endosomal maturation and intracellular trafficking pathways, playing a role in the coordination of phosphoinositide signaling on endosomal membranes.

The WDR91 Mouse mAb (Ab351136) is a monoclonal antibody produced in mouse and supplied in PBS buffer without additional carrier proteins or preservatives unless otherwise stated on the product datasheet. Researchers should consult the lot-specific Certificate of Analysis (CoA) for confirmed concentration, purity, and validated applications prior to use.

Sample Preparation

Proper sample preparation is critical for obtaining reliable results with WDR91 Mouse mAb. The following general guidelines apply; optimize conditions for your specific sample type and application:

• Cell lysates: Lyse cells in an appropriate lysis buffer (e.g., RIPA buffer) supplemented with protease inhibitor cocktail. Centrifuge at 12,000 × g for 15 minutes at 4°C to remove debris. Determine protein concentration by BCA or Bradford assay.
• Tissue lysates: Homogenize fresh or snap-frozen tissue in lysis buffer on ice. Centrifuge as above and collect the supernatant.
• Formalin-fixed paraffin-embedded (FFPE) sections: Perform antigen retrieval using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) as appropriate. Allow sections to cool before proceeding to blocking.
• Frozen sections: Fix briefly in 4% paraformaldehyde in PBS for 10–15 minutes at room temperature, then wash thoroughly with PBS.

Note: Specific protocols for each application should be optimized empirically. Refer to the product datasheet for validated applications.

Recommended Protocol Steps

The following generalized protocol is suitable as a starting point for Western blot (WB) and immunofluorescence (IF) applications using WDR91 Mouse mAb. Always refer to the product datasheet for application-specific validated dilutions.

Western Blot (WB)

1. Resolve protein samples by SDS-PAGE and transfer to PVDF or nitrocellulose membrane.
2. Block membrane with 5% non-fat milk or BSA in TBST for 1 hour at room temperature.
3. Incubate with WDR91 Mouse mAb at the recommended dilution (refer to datasheet) in blocking buffer overnight at 4°C.
4. Wash membrane 3 × 10 minutes with TBST.
5. Incubate with appropriate HRP-conjugated anti-mouse secondary antibody for 1 hour at room temperature.
6. Wash 3 × 10 minutes with TBST and detect signal using ECL substrate.

Immunofluorescence (IF)

1. Fix and permeabilize cells or tissue sections as appropriate.
2. Block with 5% normal serum in PBS for 1 hour at room temperature.
3. Incubate with WDR91 Mouse mAb at the recommended dilution overnight at 4°C.
4. Wash 3 × 5 minutes with PBS.
5. Incubate with fluorophore-conjugated anti-mouse secondary antibody for 1 hour at room temperature, protected from light.
6. Wash, counterstain nuclei with DAPI if desired, and mount coverslip.
7. Image using appropriate fluorescence microscope settings.

Note: Dilution ranges and incubation times should be optimized for each laboratory and application. Confirmed validated dilutions are provided on the product datasheet.

Troubleshooting & Optimization Tips

Use the following troubleshooting guide to address common issues encountered when using WDR91 Mouse mAb:

Always include a positive control (lysate or tissue known to express WDR91) and a negative control (primary antibody omitted) in each experiment.

Storage, Handling & Quality Notes

Storage: Store WDR91 Mouse mAb at -20°C upon receipt. For short-term use (up to 1 month), the antibody may be stored at 4°C. Avoid repeated freeze-thaw cycles, as this may reduce antibody activity and increase aggregation.

Handling: Centrifuge the vial briefly before opening to collect all liquid at the bottom. Use sterile technique to prevent microbial contamination. Dispense into single-use aliquots to minimize freeze-thaw cycles.

Formulation: This antibody is supplied in PBS (Phosphate Buffered Saline). No sodium azide or carrier protein is noted in the available product description; confirm on the lot-specific CoA before use with live-cell assays or coupling reactions.

Quality: Each lot is tested for performance consistency. Researchers are encouraged to request the Certificate of Analysis (CoA) for lot-specific performance data including validated applications, recommended dilutions, and specificity confirmation.