Recombinant Mouse BLC/BCA-1

Recombinant Mouse BLC/BCA-1 (CXCL13) is a valuable tool for research applications due to its well-defined biological activity and specific role in immune cell migration and organization. Here are key reasons to use it in your research:

1. Specific B Lymphocyte Chemoattraction

Recombinant Mouse BLC/BCA-1 is a potent and selective chemoattractant for B lymphocytes (both B-1 and B-2 subsets), but not for T lymphocytes, monocytes, or neutrophils. This specificity makes it ideal for studies focused on B cell migration, trafficking, and homing to lymphoid follicles in the spleen, lymph nodes, and Peyer’s patches.

2. Critical Role in Lymphoid Organ Development

BLC/BCA-1 signaling through its receptor CXCR5 (BLR1) is essential for the development and organization of secondary lymphoid organs. Studies in mice lacking BLR1 (CXCR5) show impaired formation of lymph nodes, Peyer’s patches, and germinal centers, highlighting the importance of this pathway in immune system architecture.

3. Use in Functional Assays

Recombinant BLC/BCA-1 is widely used in bioassays to:

• Study B cell chemotaxis and migration.
• Investigate the mechanisms of lymphoid follicle formation.
• Examine the role of CXCL13 in immune responses, inflammation, and autoimmune diseases.

4. Relevance in Disease Models

BLC/BCA-1 has been implicated in various disease contexts, including:

• Chronic pain models (e.g., CRPS-I) via CXCR5-mediated NF-κB activation and pro-inflammatory cytokine production.
• Autoimmune diseases and cancer, where CXCL13-CXCR5 signaling influences B cell autoreactivity and tumor microenvironment.

5. High Purity and Consistency

Recombinant proteins are produced under controlled conditions, ensuring high purity, batch-to-batch consistency, and reproducibility in experimental results.

6. Versatile Applications

Recombinant Mouse BLC/BCA-1 can be used in a variety of experimental setups, including:

• Cell culture studies.
• ELISA standards.
• In vivo and in vitro migration assays.
• Studies of immune cell signaling pathways.

7. Species-Specific Research

For studies in mouse models, using recombinant mouse BLC/BCA-1 ensures species-specific activity and avoids potential cross-reactivity issues that may arise with human or other species’ proteins.

In summary, Recombinant Mouse BLC/BCA-1 is essential for research involving B cell biology, lymphoid organ development, immune responses, and disease mechanisms, offering specificity, reliability, and broad applicability in immunological studies.

Yes, recombinant Mouse BLC/BCA-1 (CXCL13) can be used as a standard for quantification or calibration in ELISA assays, provided it is properly purified and quantified. This approach is widely accepted for generating standard curves in quantitative ELISA protocols.

Supporting details:

• Recombinant proteins as ELISA standards: Quantitative ELISA assays commonly use purified recombinant proteins as standards to generate calibration curves, allowing for accurate quantification of the target analyte in biological samples. The recombinant Mouse BLC/BCA-1 is suitable for this purpose if its concentration is accurately determined and it is free from interfering contaminants.

• Assay compatibility: Commercial ELISA kits for Mouse BLC/BCA-1 (CXCL13) are validated to detect both natural and recombinant forms of the protein, and standard curves generated with recombinant protein are parallel to those obtained with native protein, indicating comparable immunoreactivity and quantification accuracy.

• Preparation guidelines: When preparing standards, ensure the recombinant protein is reconstituted and diluted according to best practices, using appropriate buffers and following any lot-specific instructions to maintain consistency and reliability. The standard curve should cover the expected concentration range of your samples, typically from 0 to 1000 pg/mL, but may be extended if necessary.

• Validation: It is recommended to validate the use of your recombinant standard by performing spike-and-recovery experiments and parallelism tests to confirm that the assay quantifies both recombinant and endogenous BLC/BCA-1 equivalently in your sample matrix.

Best practices:

• Use a highly purified, well-characterized recombinant Mouse BLC/BCA-1 protein.
• Accurately determine the protein concentration using a reliable method (e.g., UV absorbance, BCA assay).
• Prepare serial dilutions for the standard curve in the same buffer/matrix as your samples.
• Validate recovery and parallelism to ensure assay accuracy.

In summary: Recombinant Mouse BLC/BCA-1 is appropriate for use as a standard in ELISA quantification, provided it is properly prepared and validated for your specific assay conditions.

Recombinant Mouse BLC/BCA-1 (CXCL13) has been validated in published research primarily for functional bioassays assessing chemotactic activity, as well as for use in ELISA, Western blot, and immunohistochemistry applications.

Key validated applications in published research include:

• Functional Bioassays:
  Recombinant Mouse BLC/BCA-1 is widely used to study its role as a chemoattractant for B lymphocytes and to investigate signaling through its receptor CXCR5. Numerous peer-reviewed studies have used it to assess B cell migration, lymphocyte trafficking, and immune cell signaling in vitro and ex vivo. These bioassays often measure chemotaxis, cell migration, and downstream signaling events such as NF-κB activation.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  The protein is used as a standard or positive control in ELISA kits designed to quantify CXCL13/BLC/BCA-1 in mouse serum, plasma, cell culture supernatants, and tissue lysates. This application is essential for biomarker studies and cytokine profiling.

• Western Blot:
  Recombinant Mouse BLC/BCA-1 is validated as a positive control for Western blotting to confirm antibody specificity and to detect endogenous or recombinant CXCL13 in biological samples.

• Immunohistochemistry (IHC):
  The protein is used in IHC protocols, both on frozen and paraffin-embedded tissue sections, to validate antibody staining and to study the localization of CXCL13 in tissues.

• Other Applications:
  Additional uses include immunoprecipitation, protein arrays, and as a blocking/neutralizing agent in functional studies to dissect the role of CXCL13 in immune responses.

Summary Table: Validated Applications for Recombinant Mouse BLC/BCA-1

In published research, the most common and well-documented use is in functional chemotaxis assays to study B cell migration and immune cell signaling, as well as in ELISA for quantification in biological samples. These applications are supported by multiple peer-reviewed studies and are considered standard in immunology and cell biology research.

To reconstitute and prepare Recombinant Mouse BLC/BCA-1 (CXCL13) protein for cell culture experiments, follow these general guidelines based on manufacturer recommendations and best practices:

Reconstitution

1. Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
2. Reconstitute the protein in sterile, distilled water or sterile PBS (phosphate-buffered saline), depending on the specific product formulation:
   • For most standard preparations, reconstitute to a concentration of 0.1–1.0 mg/mL.
   • If the product contains a carrier protein (such as BSA), reconstitute in sterile PBS with at least 0.1% BSA.
   • For carrier-free versions, reconstitute in sterile PBS alone.
3. Do not vortex the solution after reconstitution. Gently mix by pipetting up and down or by swirling the vial.

Preparation for Cell Culture

• For cell culture applications, it is generally recommended to use a formulation that includes a carrier protein (such as BSA) to enhance stability and prevent protein loss due to adsorption to surfaces.
• If using a carrier-free version, consider adding 0.1% BSA to your working buffer to improve protein stability.

Storage

• Short-term storage: The reconstituted protein can be stored at 2–8°C for up to 1 week.
• Long-term storage: For extended storage, aliquot the protein and store at –20°C to –80°C. Avoid repeated freeze-thaw cycles.
• If storing for long periods, further dilute the protein in a buffer containing 0.1% BSA and store in working aliquots.

Additional Notes

• Always use sterile technique to prevent contamination.
• Refer to the specific product datasheet for any unique instructions related to your protein preparation.

These steps will help ensure the stability and activity of Recombinant Mouse BLC/BCA-1 protein for your cell culture experiments.