Mouse IgG_2b Isotype Control [Clone MPC-11] — Purified in vivo GOLD™ Functional Grade

Clone MPC-11 is used in in vivo mouse studies as a mouse IgG2b isotype control antibody. Its primary function is to serve as a negative control to distinguish non-specific background effects from those caused by a test antibody of the same isotype.

Essential context and supporting details:

• MPC-11 has unknown binding specificity, meaning it does not bind to any known antigen in the mouse, minimizing unintended biological activity.
• In in vivo experiments, researchers administer MPC-11 alongside or instead of the antibody of interest to control for Fc receptor-mediated effects, non-specific binding, or immune activation that could arise from the antibody backbone itself rather than its antigen-specific variable region.
• The antibody is produced at high purity and very low endotoxin levels, essential for mouse in vivo work to avoid confounding inflammatory reactions.
• Typical applications include tumor models, immunotherapy studies, and antibody blockade or depletion experiments, where distinguishing specific from non-specific immune effects is critical.
• MPC-11 is also widely used in flow cytometry and immunofluorescence as an isotype control to help set gates and verify the specificity of staining.

In summary, MPC-11 acts as a non-reactive, isotype-matched control to ensure that any effects observed in vivo are due to the test antibody’s specificity rather than the IgG2b isotype itself or contaminants.

Based on the research literature, MPC-11 is commonly used alongside several other antibodies and proteins in experimental studies. Here are the key categories and specific examples:

Secondary Antibodies

Goat anti-rabbit antibodies are frequently used as secondary detection antibodies in ELISA assays involving MPC-11 cells, particularly those linked to alkaline phosphatase at dilutions of 1:5000. Additionally, secondary antibodies conjugated with horseradish peroxidase are employed in Western blotting applications at 1:5000 dilutions for visualization purposes.

Caspase Proteins for Apoptosis Studies

MPC-11 cells are commonly studied alongside several caspase proteins to investigate apoptosis pathways. The primary caspases examined include caspase-3, caspase-8, and caspase-9. These proteins are analyzed through Western blotting to determine their activation status when MPC-11 cells undergo apoptotic processes.

Control Antibodies

Control IgG antibodies are routinely used as negative controls in experiments involving MPC-11 cells, typically at concentrations of 200 µg/ml in flow cytometry and apoptosis assays. These serve as important baseline comparisons for specific antibody binding studies.

Detection Reagents

Several detection and analytical reagents are commonly paired with MPC-11 in experimental protocols:

• Propidium iodide (50 µg/ml) is used in flow cytometric analysis for cell cycle and apoptosis detection
• BCIP/NCP substrate for alkaline phosphatase detection in ELISA assays
• Vectastain ABC reagents for enhanced visualization in Western blotting applications

Comparative Cell Lines

In many studies, MPC-11 is used alongside other myeloma cell lines for comparative analysis, including SP2/0 and NS-1 cells, which are also multiple myeloma cell lines that respond similarly to polyclonal antibodies. Non-myeloma control cell lines such as CT26 (murine colon carcinoma) and B16 (murine melanoma) are also frequently included as negative controls.

The MPC-11 clone itself serves as an isotype control antibody targeting KLH (keyhole limpet hemocyanin) and is widely used in both in vivo and in vitro applications as a non-reactive control for mouse IgG2b antibodies.

Key findings from scientific literature on MPC-11 clones highlight this mouse myeloma cell line's fundamental role in immunology and cancer research, as well as notable advances in understanding immunoglobulin gene structure, class switching, and its use in therapy modeling.

1. Immunoglobulin Gene Structure and Sequencing:

• Researchers successfully cloned and sequenced cDNA corresponding to the entire variable region of the MPC-11 heavy chain, showing it contains 121 amino acids and belongs to subgroup II of mouse heavy chains.
• A crucial finding was the identification of a J (joining) segment of 16 residues that overlaps with part of the third hypervariable region; importantly, the cDNA revealed no discontinuity between the variable and constant regions, providing molecular insights into antibody gene architecture.

2. Heavy Chain Class Switching and Genetic Rearrangement:

• MPC-11 exhibits in vitro immunoglobulin heavy chain class switching, where IgG2a-producing cells can arise at high frequency from the IgG2b-producing parent line.
• DNA rearrangements on the expressed heavy chain chromosome of several MPC-11 class switch variants were mapped, confirming that the parental heavy chain variable region (VH) gene remains consistently expressed across switch variants.
• This system provides a valuable model for examining the dynamics of immunoglobulin heavy chain class switching and for mapping structural rearrangements at the molecular level.

3. Utility in Cancer Immunotherapy Research:

• The MPC-11 cell line serves as a syngeneic mouse myeloma model, compatible with BALB/c mice, and is widely used for studying anti-cancer immunotherapeutic efficacy, including immunotherapy and pharmacodynamics (PD) assessment.
• Its predictable tumor growth characteristics, either subcutaneously or via systemic injection, make it a standard model for preclinical testing of immunological interventions.

Additional Context:

• The MPC-11 system is frequently referenced in foundational studies on antibody gene rearrangement and synthesis, underpinning advances in both basic immunology and translational cancer research.
• While not a direct citation from the provided results, subsequent literature often builds on MPC-11 findings for genetically engineered antibody production and checkpoint blockade studies.

These findings collectively establish MPC-11 as a cornerpiece for molecular immunology research and a robust preclinical cancer model.

Clone MPC-11, a mouse IgG2b isotype control, is typically used as a negative control rather than a therapeutic or functional antibody, so its dosing regimen depends on its application as a control for various experimental settings rather than target activity. Published dosing regimens for MPC-11 in mouse models vary based on the context in which it is used, most commonly mirroring the dose and schedule of the test antibody it is intended to control for.

Key details on dosing regimens across mouse models:

• Standard Dose: In vivo regimens most frequently use 100–250??g per mouse per dose for isotype control antibodies such as MPC-11, often administered by intraperitoneal injection (i.p.), to match the biological exposure of experimental antibodies. Some protocols may use doses as low as 50??g or as high as 500??g per mouse, depending on the study design, mouse strain, and intended experimental power (e.g., tumor burden, organ size, or immune cell depletion).

• Dosing Frequency/Schedule:

  • Matching test antibody: Typical schedules are every 2–4 days, often for a total of 3–5 doses, closely following the dosing regimen of the test mAb (e.g., anti-PD-1, anti-CTLA-4, anti-CD4, anti-CD8). This helps ensure that mice receiving the control antibody experience the same injection schedule, potential immunogenicity, and systemic exposure.
  • Chronic or extended protocols: In models with prolonged treatment, the regimen may extend over several weeks, maintaining dose frequency to parallel experimental conditions.

• Route of Administration: Intraperitoneal (i.p.) injection is standard, though other routes (intravenous, subcutaneous) may be chosen to match the experimental antibody or model requirements.

• Mouse Model and Application Variance:

  • Tumor Models: Dosing in syngeneic or xenograft models typically aligns with the biologic targeting mAb, ranging from 100–250??g/dose for immune checkpoint inhibitor controls.
  • Depletion and Immunology Studies: For T-cell or neutrophil depletion studies, isotype controls are dosed equivalently to the depleting antibody (commonly 200–250??g per mouse, i.p., every 2–3 days).
  • In Vitro and Flow Cytometry: Doses are significantly lower (nanogram amounts per test), not directly comparable to in vivo applications.

Summary Table: MPC-11 Typical Dosing Regimens in Mouse Models

Additional Notes:

• The dosing of MPC-11 should always match both the dose and schedule of the experimental/test antibody for adequate controls.
• The strain of mouse, disease model, and experimental endpoint can influence specific regimen adjustments, though standard dosages are widely used due to their established inert profile.
• MPC-11 is used specifically as a negative control and does not have functional (depleting or blocking) activity.

References:

• : Background on MPC-11 as a standard isotype control, used as a non-reactive comparator.
• : Dosing guides for mouse mAbs establish typical ranges and frequencies for in vivo antibody use; these apply equivalently to isotype controls.
• : Manufacturing details highlight MPC-11's use in vivo, including purity, endotoxin levels, and minimal cross-reactivity.
• : Example of in vitro/flow cytometry use, where the required amount is much smaller.

If information is needed for a specific disease model or mouse strain, please clarify for more tailored data.