Target Biology: What Is AREL1?

AREL1 (Apoptosis-Resistant E3 Ubiquitin Protein Ligase 1) is a HECT-domain-containing E3 ubiquitin ligase that plays a central role in regulating programmed cell death. By ubiquitinating and targeting pro-apoptotic substrates — including SMAC/DIABLO, HtrA2/OMI, and ARTS — for proteasomal degradation, AREL1 promotes cell survival and resistance to apoptotic stimuli.

AREL1 is expressed in a variety of human tissues and has been implicated in cancer cell survival, as elevated expression may confer resistance to apoptosis-inducing therapies. Its characterization is therefore of significant interest for oncology research, cell death pathway studies, and potential therapeutic target identification.

Product Specifications

The AREL1 Mouse mAb (Ab351155) is produced in mouse and supplied as a monoclonal antibody formulated in PBS only, with no additional stabilizers or preservatives listed. Key specifications are summarized below:

• SKU: Ab351155
• Host: Mouse
• Antibody Class: Monoclonal antibody (mAb)
• Target: Human AREL1
• Formulation: PBS only
• Reference: absea#KC-2345

Researchers should store the antibody according to standard cold-chain guidelines for monoclonal antibodies. For long-term storage, aliquoting and freezing at −20°C or −80°C is recommended to avoid repeated freeze-thaw cycles. Because the formulation is PBS only, users should consider adding a carrier protein (e.g., BSA) or glycerol for long-term stability if not using the antibody immediately.

Recommended Applications & Usage Guidelines

AREL1 Mouse mAb is suitable for standard immunodetection applications. Typical uses for monoclonal antibodies of this class include:

• Western Blotting (WB): Detection of endogenous or overexpressed AREL1 protein in cell or tissue lysates. Recommended to optimize primary antibody concentration empirically.
• Immunoprecipitation (IP): Pulldown of AREL1 and its associated protein complexes for downstream analysis.
• Immunofluorescence (IF): Subcellular localization studies of AREL1 in fixed cells.
• Immunohistochemistry (IHC): Detection of AREL1 expression in formalin-fixed, paraffin-embedded (FFPE) tissue sections.

Note: Specific validated applications and working dilutions are not detailed in the available source material. Users should perform titration experiments and consult the product datasheet for confirmed application data.

General Protocol for Western Blotting with AREL1 Mouse mAb

The following is a general guideline for using AREL1 Mouse mAb in Western blotting. Optimize conditions according to your specific cell/tissue system.

1. Sample Preparation: Lyse cells or homogenize tissue in RIPA buffer supplemented with protease inhibitor cocktail. Determine protein concentration by BCA or Bradford assay. Load 20–50 µg total protein per lane.
2. SDS-PAGE: Resolve proteins on a 8–12% polyacrylamide gel under reducing conditions, then transfer to PVDF or nitrocellulose membrane.
3. Blocking: Block membrane for 1 hour at room temperature in 5% non-fat dry milk or 5% BSA in TBST.
4. Primary Antibody Incubation: Incubate with AREL1 Mouse mAb at an empirically determined dilution (suggested starting range: 1:500–1:2000) overnight at 4°C with gentle agitation.
5. Washing: Wash 3 × 10 minutes in TBST.
6. Secondary Antibody: Incubate with an HRP-conjugated anti-mouse IgG secondary antibody (1:5000–1:10000) for 1 hour at room temperature.
7. Detection: Wash 3 × 10 minutes in TBST, then develop using ECL substrate and image on film or a digital imager.

Expected band size for AREL1: approximately 100 kDa (verify against the product datasheet, as the precise molecular weight should be confirmed).