Recombinant Human CHMP4BP1 Protein: Protocol & Usage Guide

Recombinant Human CHMP4BP1 Protein is a research-grade recombinant protein designed for studies involving the ESCRT machinery, multivesicular body biogenesis, membrane remodeling, and cytokinesis-related biological processes.

Product Name

Recombinant Human CHMP4BP1 Protein

SKU

rp376263

Species

Human (Homo sapiens)

Protein Family

CHMP (Charged Multivesicular Body Protein) family, ESCRT-III associated

Primary Application

Biochemical assays, protein interaction studies, ESCRT pathway research

Storage Recommendation

Store at -80°C; avoid repeated freeze-thaw cycles

Reference Source

absea #PP-9870

Overview of CHMP4BP1 and Biological Significance

CHMP4BP1 (Charged Multivesicular Body Protein 4B-binding Protein 1) is a protein that interacts with CHMP4B, a core component of the ESCRT-III (Endosomal Sorting Complexes Required for Transport III) machinery. The ESCRT pathway is essential for a range of membrane remodeling events including multivesicular body (MVB) biogenesis, viral budding, and cytokinesis abscission.

CHMP4BP1 has been implicated in the regulation of CHMP4B localization and function, influencing ESCRT-III assembly dynamics at the midbody during cytokinesis. Research into CHMP4BP1 contributes to understanding how ESCRT complexes are spatially and temporally regulated in dividing cells.

Recombinant Human CHMP4BP1 Protein (SKU: rp376263) provides researchers with a well-defined reagent for in vitro biochemical studies, binding assays, and mechanistic investigations of the ESCRT-III regulatory network.

Reconstitution and Storage Protocol

Materials Required:

• Recombinant Human CHMP4BP1 Protein (rp376263)
• Sterile deionized water or manufacturer-recommended reconstitution buffer
• Microcentrifuge tubes (low-protein-binding recommended)
• Ice or cooling block

Reconstitution Steps:

1. Briefly centrifuge the vial at low speed (approximately 200–500 × g for 5–10 seconds) before opening to collect lyophilized material at the bottom of the vial.
2. Add the appropriate volume of sterile reconstitution buffer (refer to the product datasheet for the specific recommended buffer and volume) dropwise to the vial.
3. Gently swirl or pipette the solution to dissolve the protein. Do not vortex, as vigorous agitation may cause protein denaturation.
4. Allow the solution to sit on ice for 5–10 minutes to ensure complete dissolution.
5. Aliquot the reconstituted protein into single-use volumes to minimize freeze-thaw cycles.

Storage Conditions:

• Store lyophilized protein at -20°C or -80°C as specified on the product datasheet.
• Store reconstituted protein at -80°C for long-term storage.
• Avoid repeated freeze-thaw cycles, which may reduce protein activity and integrity.
• Short-term storage (up to 1 week) may be performed at 4°C if a stabilizing carrier protein such as BSA is included.

Recommended Applications and Experimental Use

Recombinant Human CHMP4BP1 Protein is suitable for a variety of in vitro research applications. Researchers should validate the protein for each specific application and optimize experimental conditions accordingly.

Recommended Applications Include:

• Protein–Protein Interaction Studies: Use CHMP4BP1 protein in pull-down assays or co-immunoprecipitation setups to characterize its interaction with CHMP4B and other ESCRT-III components.
• Biochemical Activity Assays: Investigate regulatory roles of CHMP4BP1 in ESCRT-III assembly or disassembly in cell-free reconstitution systems.
• Structural Studies: Use purified recombinant protein as input material for biophysical analyses such as size-exclusion chromatography, dynamic light scattering, or cryo-electron microscopy sample preparation.
• Antibody Development and Validation: Use as an antigen standard for ELISA calibration or Western blot positive controls.

Working Concentration: Optimal working concentrations should be determined empirically for each assay system. Refer to the product datasheet for suggested starting concentrations.

Important Note: This product is for research use only (RUO). It is not intended for diagnostic, therapeutic, or clinical use.

Quality Control and Handling Precautions

Quality Control:

Each lot of Recombinant Human CHMP4BP1 Protein is subject to quality control testing prior to release. Typical QC analyses include:

• SDS-PAGE analysis to confirm molecular weight and purity
• Bioactivity testing where applicable
• Endotoxin testing by LAL (Limulus Amebocyte Lysate) assay

Specific QC data, including purity percentage, endotoxin levels, and lot-specific results, are provided on the product Certificate of Analysis (CoA). Request the CoA for your specific lot number from Aladdin Scientific.

Handling Precautions:

• Always work on ice when handling reconstituted protein to maintain stability.
• Use low-protein-binding pipette tips and tubes to minimize protein loss due to adsorption.
• Centrifuge briefly before use if any precipitation is visible.
• Do not use protein that shows signs of irreversible aggregation or significant discoloration.
• Standard laboratory safety precautions should be followed; refer to the Safety Data Sheet (SDS) for this product.

Frequently asked questions

What is CHMP4BP1 and why is it relevant to ESCRT research?

CHMP4BP1 is a binding partner of CHMP4B, a key component of the ESCRT-III complex involved in membrane remodeling, multivesicular body biogenesis, viral budding, and cytokinesis abscission. Studying CHMP4BP1 helps elucidate how ESCRT-III assembly and function are regulated in space and time within cells.

How should I reconstitute Recombinant Human CHMP4BP1 Protein?

Briefly centrifuge the vial before opening, then add the recommended volume of sterile reconstitution buffer dropwise. Gently swirl to dissolve—do not vortex. Allow the solution to sit on ice for 5–10 minutes, then aliquot into single-use volumes and store at -80°C. Always refer to the product datasheet for buffer specifications and recommended volumes.

How many freeze-thaw cycles can Recombinant Human CHMP4BP1 Protein withstand?

Repeated freeze-thaw cycles are not recommended as they can reduce protein activity and structural integrity. It is best practice to aliquot the reconstituted protein into single-use volumes immediately after reconstitution to avoid repeated freeze-thaw cycles.

Is Recombinant Human CHMP4BP1 Protein suitable for in vivo or clinical use?

No. This product is for research use only (RUO) and is not approved or intended for diagnostic, therapeutic, or clinical applications. Researchers should adhere to all applicable institutional and regulatory guidelines when using this reagent.

Where can I obtain a Certificate of Analysis (CoA) for my specific lot?

Contact Aladdin Scientific's customer service team with your lot number and order information to request the Certificate of Analysis. The CoA provides lot-specific quality control data including purity, endotoxin levels, and activity results.

References

1. absea Product Reference: PP-9870. Recombinant Human CHMP4BP1 Protein.
2. Hurley JH. ESCRTs are everywhere. EMBO J. 2015;34(19):2398-2407. doi:10.15252/embj.201592484
3. Morita E, Sundquist WI. Retrovirus budding. Annu Rev Cell Dev Biol. 2004;20:395-425. doi:10.1146/annurev.cellbio.20.010403.102350

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