Target Background: What Is RNF113A?

RNF113A (Ring Finger Protein 113A) is a RING domain-containing protein that functions as part of the cellular ubiquitination machinery. RING finger domain proteins typically act as E3 ubiquitin ligases, facilitating the transfer of ubiquitin from an E2 ubiquitin-conjugating enzyme to specific substrate proteins, thereby regulating protein stability, localization, and activity.

RNF113A has been studied in the context of RNA splicing and DNA damage response pathways. It is associated with the spliceosome and has been implicated in the regulation of gene expression at the post-transcriptional level. Research interest in RNF113A has grown due to its potential involvement in cancer biology and cellular stress responses.

Reliable detection of RNF113A protein expression is essential for researchers investigating ubiquitin-proteasome pathway components, RNA processing complexes, or disease-associated alterations in protein homeostasis.

Antibody Specifications & Formulation Guidance

The RNF113A Mouse mAb (Ab351054) is a monoclonal antibody produced in mouse. The product is supplied in PBS (phosphate-buffered saline) only, without additional carrier proteins such as BSA or glycerol as stabilizers, based on the available product description referencing Absea clone KC-27539.

Key formulation considerations for PBS-only antibodies:

• Protein concentration awareness: PBS-only formulations without stabilizing proteins may be more susceptible to freeze-thaw degradation. Aliquoting upon receipt is strongly recommended to minimize repeated freeze-thaw cycles.
• Storage temperature: Store at -20°C for long-term storage. Avoid repeated freeze-thaw cycles. Short-term storage (up to 1 week) may be performed at 4°C.
• Do not dilute before aliquoting: Prepare working aliquots at the stock concentration, then dilute only immediately before use.

Researchers should confirm the exact concentration, isotype, and application-validated data by consulting the full product datasheet available from Aladdin Scientific for Ab351054.

Recommended Protocols & Application Guidelines

Monoclonal antibodies targeting RING finger proteins such as RNF113A are commonly employed in the following immunological techniques. Specific validated applications for Ab351054 should be confirmed on the product datasheet, as performance can vary by lot and application.

Western Blot (WB)

Western blotting is one of the most common applications for anti-RNF113A antibodies. Typical starting conditions include:

• Primary antibody dilution: 1:500–1:2000 (optimize empirically)
• Blocking: 5% non-fat dry milk or 3–5% BSA in TBST, 1 hour at room temperature
• Incubation: Overnight at 4°C with gentle rocking
• Expected molecular weight of RNF113A: approximately 50–60 kDa (verify against datasheet)

Immunoprecipitation (IP)

For IP applications, use 1–5 µg of antibody per 500 µg of total cell lysate. Pre-clear lysates with protein A/G beads before adding the primary antibody to reduce non-specific binding.

Immunofluorescence (IF) / Immunocytochemistry (ICC)

For IF/ICC, fix cells in 4% paraformaldehyde, permeabilize with 0.1–0.2% Triton X-100, and block with 5% normal serum. Use a validated secondary anti-mouse antibody conjugated to an appropriate fluorophore.

Note: All dilutions and incubation conditions should be optimized for each specific experimental system. Consult the Ab351054 datasheet for application-specific validation data.

Storage, Handling & Stability Best Practices

Proper storage and handling are critical to maintaining the activity and specificity of monoclonal antibodies. For RNF113A Mouse mAb (Ab351054) supplied in PBS only, follow these guidelines:

• Upon receipt: Immediately inspect for visible particulates or color changes. Centrifuge briefly if necessary to collect all liquid.
• Aliquoting: Divide into single-use aliquots to prevent repeated freeze-thaw. Label each aliquot with date, concentration, and lot number.
• Long-term storage: -20°C (or -80°C for extended storage). Do not store in a frost-free freezer that undergoes automatic defrost cycles.
• Working stock: After thawing, keep at 4°C and use within 1–2 weeks. Do not refreeze thawed aliquots if avoidable.
• Avoid: Repeated freeze-thaw cycles, exposure to light (if using fluorophore-conjugated formats), and contamination with foreign proteins or microorganisms.
• Carrier protein addition: If long-term stability at low concentrations is a concern, addition of BSA (to 0.1%) or glycerol (to 10–50%) may be considered, but this should be validated against downstream applications.

Always consult the product datasheet and Aladdin Scientific technical support for lot-specific stability data.

Troubleshooting Common Issues

Even with high-quality monoclonal antibodies, experimental issues can arise. Below are common problems and recommended corrective actions for RNF113A Mouse mAb experiments:

Contact Aladdin Scientific technical support if issues persist after standard troubleshooting steps. Provide lot number, protocol details, and images when seeking assistance.