PON3 Mouse mAb Protocol: Applications & Usage Guide

PON3 Mouse mAb is a monoclonal antibody raised in mouse against human PON3 (Paraoxonase 3), supplied in PBS and validated for immunological detection of PON3 protein in research applications.

Product Name

PON3 Mouse mAb

SKU

Ab350645

Host Species

Mouse

Antibody Type

Monoclonal Antibody (mAb)

Formulation

PBS only

Target Protein

PON3 (Paraoxonase 3)

Reference Source

absea #KC-28343

Overview of PON3 and the Monoclonal Antibody

Paraoxonase 3 (PON3) is a member of the paraoxonase family of enzymes, which includes PON1, PON2, and PON3. PON3 is a calcium-dependent lactonase primarily associated with high-density lipoprotein (HDL) and plays a role in antioxidant defense and lipid metabolism. Dysregulation of PON3 has been implicated in cardiovascular disease, oxidative stress, and metabolic disorders.

PON3 Mouse mAb (Ab350645) is a monoclonal antibody raised in mouse specifically targeting PON3. It is supplied in PBS without additional preservatives or carrier proteins, making it suitable for a variety of downstream immunological applications. This product is referenced as absea #KC-28343.

Recommended Protocol

Sample Preparation: Prepare cell lysates or tissue homogenates in an appropriate lysis buffer supplemented with protease inhibitors. Determine protein concentration using a standard assay (e.g., BCA or Bradford) prior to use.

Western Blot (WB):

1. Resolve protein samples by SDS-PAGE and transfer to a PVDF or nitrocellulose membrane.
2. Block the membrane with 5% non-fat milk or BSA in TBST for 1 hour at room temperature.
3. Incubate with PON3 Mouse mAb at the recommended working dilution (see manufacturer's datasheet for verified dilution range) overnight at 4°C.
4. Wash three times with TBST (5 minutes each).
5. Incubate with an appropriate anti-mouse HRP-conjugated secondary antibody for 1 hour at room temperature.
6. Wash three times with TBST and detect signal using a chemiluminescent substrate.

ELISA:

1. Coat ELISA plate wells with antigen at 1–4 µg/mL in carbonate-bicarbonate coating buffer overnight at 4°C.
2. Block with 1% BSA in PBS for 1–2 hours at room temperature.
3. Add PON3 Mouse mAb at the appropriate dilution and incubate for 1–2 hours at room temperature.
4. Wash and add HRP-conjugated anti-mouse secondary antibody. Incubate for 1 hour.
5. Develop with TMB substrate, stop with stop solution, and read absorbance at 450 nm.

Note: Optimal working dilutions should be determined empirically for each application and experimental system. Refer to the product datasheet for verified conditions.

Storage and Handling

PON3 Mouse mAb (Ab350645) is supplied in PBS only. To ensure optimal antibody performance, observe the following storage and handling guidelines:

• Short-term storage (up to 1 month): Store at 4°C.
• Long-term storage: Aliquot the antibody to avoid repeated freeze-thaw cycles and store at −20°C or below. Adding a carrier protein such as 0.1% BSA may help stabilize the antibody for long-term storage if no BSA is already present.
• Avoid: Repeated freeze-thaw cycles, exposure to direct light, and contamination.
• Centrifuge vial briefly before opening to collect contents at the bottom.
• Keep on ice during experimental use.

Note: Specific storage recommendations from the manufacturer's datasheet take precedence. Always refer to the product insert for the most current guidance.

Troubleshooting

If unexpected results are obtained when using PON3 Mouse mAb, consider the following troubleshooting steps:

Issue	Possible Cause	Suggested Action
No signal detected	Antibody concentration too low; antigen not expressed in sample	Increase antibody concentration; verify PON3 expression in your cell/tissue model; use a positive control lysate
High background	Insufficient blocking; antibody concentration too high	Extend blocking time; reduce primary antibody concentration; increase wash stringency
Non-specific bands (WB)	Cross-reactivity; degraded sample	Optimize antibody dilution; use fresh samples with protease inhibitors; add reducing agent if appropriate
Weak signal	Over-washing; poor transfer efficiency	Reduce wash duration; verify transfer using a pre-stained marker or Ponceau S staining

Frequently asked questions

What is the formulation of PON3 Mouse mAb (Ab350645)?

PON3 Mouse mAb (Ab350645) is supplied in PBS only, without additional stabilizers, preservatives, or carrier proteins. Researchers requiring additional stabilization for long-term storage may consider adding BSA or glycerol, but should validate any additives do not interfere with downstream applications.

Which applications is PON3 Mouse mAb validated for?

Based on the available product information, PON3 Mouse mAb is suitable for immunological detection of PON3. Common applications for mouse monoclonal antibodies of this type include Western blot (WB) and ELISA. Researchers should consult the official product datasheet for a complete and verified list of validated applications and corresponding dilution ranges.

How should PON3 Mouse mAb be stored to maintain activity?

For short-term use (up to 1 month), store at 4°C. For long-term storage, aliquot the antibody into single-use volumes and store at −20°C or below to avoid repeated freeze-thaw cycles, which can degrade antibody integrity. Keep away from direct light and contamination.

What is PON3 and why is it an important research target?

PON3 (Paraoxonase 3) is a calcium-dependent lactonase associated with HDL particles and plays a protective role in oxidative stress and lipid peroxidation. It is of significant interest in research on cardiovascular disease, metabolic syndrome, and antioxidant defense mechanisms. An antibody specific to PON3 enables precise detection and quantification of this enzyme in biological samples.

Is there a recommended positive control for experiments using PON3 Mouse mAb?

Cell lines or tissues known to express PON3, such as hepatic (liver) cell lines, are commonly used as positive controls for PON3 antibody validation. Researchers should verify PON3 expression levels in their specific model system prior to experimental use. Consult the product datasheet for any manufacturer-recommended controls.

References

1. absea KC-28343 product reference. Absea Biotechnology Ltd. PON3 Monoclonal Antibody.
2. Draganov DI, Teiber JF, Speelman A, Osawa Y, Sunahara R, La Du BN. Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities. J Lipid Res. 2005;46(6):1239-1247. doi:10.1194/jlr.M400511-JLR200

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