PSMC3IP Antibody Usage Guidelines | Research Protocol

PSMC3IP Antibody is a research-grade immunoreagent designed to detect PSMC3IP, a key regulatory protein involved in proteasome assembly and DNA damage response pathways.

Catalog Number

Ab350658

Product Name

PSMC3IP Antibody

Formulation

PBS only (no carrier protein or preservative added)

Reference Clone / Source Code

absea#OC-683

Recommended Storage

Store at -20 °C; avoid repeated freeze-thaw cycles

Target Protein

PSMC3IP (Proteasome 26S Subunit, ATPase Interacting Protein)

Background: PSMC3IP and Its Biological Role

PSMC3IP (Proteasome 26S Subunit, ATPase 3-Interacting Protein), also known as HOP2 or TBPIP, is a nuclear protein that plays dual roles in mammalian cells. It functions as a co-activator of the 26S proteasome regulatory ATPase PSMC3, contributing to ubiquitin-mediated protein degradation. Additionally, PSMC3IP participates in homologous recombination-based DNA repair, where it forms a complex with MND1 to facilitate RAD51- and DMC1-mediated strand invasion during meiosis and mitotic DNA damage repair.

Dysregulation of PSMC3IP has been implicated in ovarian dysfunction and premature ovarian failure (POF), making it a target of interest in reproductive biology and oncology research. Antibodies against PSMC3IP enable researchers to study its expression levels, subcellular localization, and protein–protein interactions across diverse experimental models.

Product Specifications and Formulation

The PSMC3IP Antibody (Cat. No. Ab350658) is supplied in phosphate-buffered saline (PBS) without the addition of glycerol, BSA, or sodium azide preservative. The absence of carrier proteins and preservatives in the PBS-only formulation makes this antibody particularly suitable for applications where exogenous proteins or azide interference must be minimized, such as conjugation reactions, proximity ligation assays, or certain enzyme-linked immunoassays.

Key Specifications:

• Catalog No.: Ab350658
• Source Code: absea#OC-683
• Buffer: PBS only
• Recommended Storage: -20 °C (long-term); 4 °C (short-term, up to 1 month)
• Avoid: Repeated freeze-thaw cycles — aliquot upon receipt if multiple uses are anticipated

Note: Specific host species, clonality, isotype, and validated applications are not detailed in the current source material. Please contact Aladdin Scientific technical support or consult the product datasheet for complete specifications.

General Protocol Guidelines for PSMC3IP Antibody Use

The following guidelines represent general best practices for antibody-based detection of intracellular nuclear targets such as PSMC3IP. Users should validate optimal conditions empirically for each application and sample type.

Western Blotting (WB)

1. Prepare whole-cell or nuclear lysates under denaturing conditions (RIPA or equivalent lysis buffer with protease inhibitor cocktail).
2. Separate proteins by SDS-PAGE and transfer to PVDF or nitrocellulose membrane.
3. Block membrane with 5% non-fat dry milk or 5% BSA in TBST for 1 hour at room temperature.
4. Incubate with PSMC3IP Antibody at the empirically determined dilution (typical starting range: 1:500–1:2000) in blocking buffer overnight at 4 °C.
5. Wash 3× with TBST (10 minutes each), then incubate with appropriate HRP-conjugated secondary antibody for 1 hour at room temperature.
6. Detect signal using chemiluminescent substrate and image on film or digital imaging system.

Immunofluorescence (IF) / Immunocytochemistry (ICC)

1. Fix cells with 4% paraformaldehyde (10 minutes, room temperature) and permeabilize with 0.1–0.3% Triton X-100.
2. Block with 5% normal serum (matched to secondary antibody host) for 30–60 minutes.
3. Apply PSMC3IP Antibody at recommended dilution and incubate overnight at 4 °C.
4. Wash and detect with fluorophore-conjugated secondary antibody. Counterstain nuclei with DAPI.

All dilutions and incubation times should be optimized by the end user. Positive and negative controls are strongly recommended in every experiment.

Troubleshooting Common Issues

If unexpected results are obtained when using the PSMC3IP Antibody, consider the following troubleshooting steps:

Problem	Possible Cause	Suggested Action
No signal detected	Antibody degraded due to improper storage or repeated freeze-thaw	Aliquot antibody upon receipt; store at -20 °C; use fresh aliquot
High background	Insufficient blocking or antibody concentration too high	Extend blocking time; optimize antibody dilution; increase wash stringency
Non-specific bands (WB)	Cross-reactivity or overloading	Reduce protein load; titrate antibody; consult datasheet for expected molecular weight
Weak nuclear signal (IF)	Inadequate permeabilization	Increase Triton X-100 concentration or extend permeabilization time
Signal loss after long storage	PBS-only formulation lacks stabilizing agents	Add glycerol (50% v/v final) for -20 °C storage; aliquot to minimize freeze-thaw

Ordering Information and Technical Support

The PSMC3IP Antibody is available exclusively through Aladdin Scientific. Place orders via the Aladdin Scientific website or contact your regional sales representative.

• Catalog No.: Ab350658
• Product: PSMC3IP Antibody
• Formulation: PBS only
• Source Reference: absea#OC-683

For technical inquiries regarding optimal working dilutions, validated applications, species cross-reactivity, or custom conjugation services, please contact Aladdin Scientific Technical Support. Our team can provide application-specific guidance and access to the latest product datasheet.

Note for Procurement: This product is intended for research use only (RUO) and is not approved for diagnostic or therapeutic use in humans or animals.

Frequently asked questions

What buffer is the PSMC3IP Antibody (Ab350658) supplied in?

The PSMC3IP Antibody is supplied in PBS (phosphate-buffered saline) only, without glycerol, BSA, or sodium azide. Users requiring long-term storage stability may consider adding glycerol to a final concentration of 50% v/v before storing at -20 °C.

How should I store the PSMC3IP Antibody to maintain its activity?

Store the antibody at -20 °C for long-term use or at 4 °C for short-term use (up to one month). Because the formulation is PBS only and lacks stabilizing agents such as glycerol, it is strongly recommended to aliquot the antibody into single-use volumes upon receipt to avoid repeated freeze-thaw cycles, which can cause protein aggregation and loss of immunoreactivity.

What applications is the PSMC3IP Antibody suitable for?

General best-practice protocols for nuclear protein antibodies support use in applications such as Western blotting (WB), immunofluorescence (IF), and immunocytochemistry (ICC). However, the specific validated applications for Ab350658 are not fully detailed in the current source material. Please refer to the official product datasheet or contact Aladdin Scientific Technical Support for confirmed application data.

Can the PBS-only formulation of PSMC3IP Antibody be used directly in conjugation reactions?

Yes, the absence of BSA, glycerol, and sodium azide in the PBS-only formulation makes the PSMC3IP Antibody particularly amenable to direct chemical conjugation (e.g., with fluorescent dyes, biotin, or enzymes), since these additives can interfere with conjugation chemistry. However, users should verify antibody concentration and purity prior to conjugation and follow the conjugation kit manufacturer's instructions.

What is the biological significance of PSMC3IP, and why is it a research target?

PSMC3IP is involved in two critical cellular processes: regulation of the 26S proteasome via interaction with the ATPase subunit PSMC3, and homologous recombination DNA repair through its MND1-interacting complex that facilitates RAD51/DMC1 strand invasion. Mutations in PSMC3IP have been linked to premature ovarian failure, and its role in DNA repair makes it relevant to cancer biology and fertility research. These diverse functions make PSMC3IP a valuable target for mechanistic studies.

References

1. Aladdin Scientific Product Datasheet: PSMC3IP Antibody, Cat. No. Ab350658 (absea#OC-683). Aladdin Scientific, internal product documentation.
2. Petukhova GV, et al. The Hop2 and Mnd1 proteins act in concert with Rad51 and Dmc1 in meiotic recombination. Nature Structural & Molecular Biology, 2005. [General reference for PSMC3IP/HOP2 biology — specific details require verification.]
3. Zangen D, et al. XX ovarian dysgenesis is caused by a PSMC3IP/HOP2 mutation that abolishes coactivation of estrogen-driven transcription. American Journal of Human Genetics, 2011. [Reference for PSMC3IP and premature ovarian failure — specific details require verification.]

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