PELI3 Mouse mAb: Selection, Application & Protocol Guide

PELI3 Mouse mAb is a mouse-derived monoclonal antibody specifically targeting the PELI3 (Pellino E3 Ubiquitin Protein Ligase 3) protein, formulated in PBS and designed to deliver consistent, reproducible performance across key immunoassay applications.

Product Name

PELI3 Mouse mAb

SKU

Ab350624

Clone / Source Reference

KC-1370 (absea)

Formulation

PBS only

Host Species

Mouse

Antibody Type

Monoclonal Antibody (mAb)

Target Protein

PELI3 (Pellino E3 Ubiquitin Protein Ligase 3)

Background: PELI3 Biology and Significance

PELI3, also known as Pellino E3 Ubiquitin Protein Ligase 3, is a member of the Pellino family of E3 ubiquitin ligases. These proteins play essential roles in innate immune signalling pathways, particularly in Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) cascades, by catalysing the ubiquitination of key signalling intermediates such as IRAK1 and IRAK4.

Dysregulation of PELI3 has been implicated in inflammatory diseases, viral pathogenesis, and cancer biology, making it a relevant target for both basic research and translational studies. Monoclonal antibodies against PELI3 are critical tools for detecting and quantifying this protein in complex biological samples.

Product Description and Formulation Details

Aladdin Scientific's PELI3 Mouse mAb (SKU: Ab350624) is a monoclonal antibody produced in mouse, derived from the absea clone KC-1370. The antibody is supplied in phosphate-buffered saline (PBS) without additional stabilisers or preservatives, making it compatible with a range of downstream conjugation and labelling workflows.

Because the formulation contains PBS only, researchers should consider adding a carrier protein (e.g., 0.1–1% BSA) and a preservative (e.g., 0.02% sodium azide) for long-term storage if the antibody is to be aliquoted and stored at –20 °C. Single-use aliquots are strongly recommended to avoid repeated freeze–thaw cycles that may compromise antibody integrity.

Recommended Applications and Protocol Guidelines

While specific validated applications for Ab350624 should be confirmed with current Aladdin Scientific product documentation, PELI3 monoclonal antibodies of similar class are generally applied in:

• Western Blot (WB): Dilute antibody in blocking buffer (e.g., 5% non-fat milk or BSA in TBST). Typical primary antibody incubation is overnight at 4 °C. Use an appropriate HRP-conjugated anti-mouse secondary antibody for detection.
• Immunohistochemistry (IHC): Apply antigen retrieval (heat-induced, citrate buffer pH 6.0 is commonly used). Incubate primary antibody at optimised dilution; use an anti-mouse HRP-polymer detection system. Counterstain with haematoxylin.
• Immunofluorescence (IF): Fix and permeabilise cells appropriately before incubation with the primary antibody. Use a fluorophore-conjugated anti-mouse secondary antibody.

Note: Optimal working dilutions must be determined empirically for each application and sample type. Researchers should run appropriate positive and negative controls in all experiments.

Storage, Handling, and Quality Considerations

Proper storage and handling are essential to maintaining the specificity and sensitivity of PELI3 Mouse mAb. Follow these general guidelines:

• Short-term storage (up to 1 month): Store at 4 °C in PBS. Add 0.02% sodium azide if microbial contamination is a concern.
• Long-term storage: Aliquot and store at –20 °C or –80 °C. Avoid repeated freeze–thaw cycles.
• Do not dilute the stock antibody until immediately before use.
• Centrifuge the vial briefly before opening to collect contents at the bottom.
• Always use appropriate personal protective equipment (PPE) and follow institutional biosafety guidelines when handling antibody preparations.

As the formulation is PBS only, no carrier protein or glycerol is present. This may make the antibody more susceptible to activity loss upon prolonged storage at 4 °C; single-use aliquots at –20 °C are therefore preferred for most laboratories.

Troubleshooting Common Issues

If unexpected results are obtained with PELI3 Mouse mAb, consider the following troubleshooting steps:

• No signal / weak signal: Verify that PELI3 is expressed in the cell line or tissue used. Check antibody concentration and incubation conditions. Ensure secondary antibody is compatible with mouse primary antibodies.
• High background: Increase blocking time or blocking agent concentration. Reduce primary or secondary antibody concentration. Increase wash stringency (number of washes or detergent concentration).
• Non-specific bands (WB): Optimise antibody dilution. Confirm sample preparation conditions, including lysis buffer composition and protease inhibitor use.
• Loss of signal over time: Check storage conditions. Repeated freeze–thaw cycles degrade mAb performance; ensure single-use aliquots are used.

For application-specific technical support, contact Aladdin Scientific's technical services team.

Frequently asked questions

What is PELI3 and why is it a relevant research target?

PELI3 (Pellino E3 Ubiquitin Protein Ligase 3) is an E3 ubiquitin ligase involved in innate immune signalling, particularly downstream of TLR and IL-1R pathways. Its role in regulating inflammation and its association with disease states such as cancer and inflammatory disorders make it a target of significant research interest.

What applications is PELI3 Mouse mAb (Ab350624) suitable for?

PELI3 Mouse mAb is expected to be applicable in common immunoassay formats including Western Blot (WB), Immunohistochemistry (IHC), and Immunofluorescence (IF). Researchers should verify validated applications in the current Aladdin Scientific product datasheet and optimise conditions empirically for their specific samples.

Why is the antibody formulated in PBS only, and does this affect storage?

The PBS-only formulation provides flexibility for downstream conjugation and labelling without interference from carrier proteins or glycerol. However, it means the antibody may be less stable during prolonged storage at 4 °C compared to formulations containing stabilisers. Single-use aliquots stored at –20 °C are recommended to preserve activity.

How should I determine the optimal working dilution for PELI3 Mouse mAb?

Optimal working dilutions must be determined empirically for each application and sample type. Start with dilution ranges recommended in the product datasheet (if provided), run a titration experiment with appropriate positive controls expressing PELI3, and adjust based on signal-to-noise ratio.

What secondary antibody should I use with PELI3 Mouse mAb?

Since PELI3 Mouse mAb is a mouse-derived monoclonal antibody, an anti-mouse secondary antibody conjugated to the appropriate detection label (e.g., HRP for WB and IHC, or a fluorophore for IF) should be used. Ensure the secondary antibody is validated for use with your detection system.

References

1. Aladdin Scientific Product Page: PELI3 Mouse mAb, SKU Ab350624. Absea clone KC-1370, PBS formulation.
2. Strelow A, et al. Pellino proteins, IRAK1 and IRAK4 ubiquitination in TLR signalling — general review (textbook-level knowledge; specific citation not provided).

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.