Anti-Mouse MAdCAM-1 (MECA-367) – Purified in vivo GOLD^TM Functional Grade

The MECA-367 clone is an anti-mouse MAdCAM-1 monoclonal antibody widely used in in vivo mouse studies to block the interaction between MAdCAM-1 (mucosal addressin cell adhesion molecule-1) and its ligand on lymphocytes, thus inhibiting lymphocyte homing to mucosal tissues.

In vivo, administration of MECA-367 is used to:

• Neutralize MAdCAM-1, thereby preventing lymphocytes (particularly ?4?7^+^ and LPAM-1^+^ T cells) from tethering, rolling, and homing to gut-associated lymphoid tissues such as Peyer’s patches and mesenteric lymph nodes.
• Reduce T cell-mediated inflammation in mouse models of gastrointestinal diseases and inflammatory bowel disease.
• Pharmacodynamically, MECA-367 administration causes a dose-dependent increase in circulating ?7^+^ memory T cells (because these cells are prevented from migrating into tissues expressing MAdCAM-1), which can be quantitated by flow cytometry after IV dosing (e.g., tail vein injection at various microgram/kilogram body weights).
• MECA-367 is often used as a surrogate for anti-human MAdCAM-1 antibodies (such as PF-00547659) in preclinical mouse models to test mechanisms, biomarkers, and dosing strategies for compounds that block leukocyte homing.

Typical in vivo protocols include:

• IV injection (most commonly via the tail vein) at doses between 0.1 and 3 mg/kg.
• Blood sample collection several days after administration (usually day 3-7), followed by analysis of lymphocyte subsets via flow cytometry.
• Sometimes, MECA-367 is used to ameliorate or probe disease pathogenesis in models of colitis, diabetes, or tissue-specific inflammation by interfering with immune cell trafficking.

Key points:

• Mechanism: Blocks lymphocyte entry into mucosal tissues by inhibiting MAdCAM-1 function.
• Readouts: Increases in circulating mucosal-homing T cell populations; reduced tissue inflammation, particularly in gut-associated tissues.
• Applications: In vivo MAdCAM-1 neutralization, mechanistic studies of leukocyte trafficking, and disease attenuation in inflammatory models.

MECA-367 is thus an established tool antibody for in vivo mouse research on immune cell migration and mucosal immunology.

The correct storage temperature for sterile packaged clone MECA-367 is 2–8°C for short-term storage (up to one month), undiluted and protected from light; for longer-term storage, aliquot and store at ? –20°C or ? –70°C, depending on manufacturer guidance.

• For immediate use or within 4 weeks, store undiluted at 2–8°C (refrigerator grade, not frozen).
• For long-term storage, aseptically aliquot and store at –20°C to –70°C (manual defrost freezer). Avoid repeated freeze-thaw cycles.

Additional details:

• Do not freeze for routine storage at 2–8°C; freezing is only advised for long-term storage after proper aliquoting.
• Always use manual defrost freezers for long-term low-temperature storage.
• Store solutions undiluted and protected from light for optimal antibody stability.

This recommendation is consistent across major antibody suppliers and package inserts for MECA-367. For unique formulations or lot-specific requirements, always consult the product datasheet accompanying your purchase.

Commonly used antibodies or proteins in the literature with MECA-367, an anti-MAdCAM-1 monoclonal antibody, include isotype controls such as rat IgG2a? (e.g., R35–95), and other adhesion molecules like PF-00547659 (an anti-human MAdCAM-1 antibody), VCAM-1, and proteins involved in lymphocyte homing such as ?4?7 (LPAM-1) and CD62L.

Key proteins and antibodies typically used with or in comparison to MECA-367:

• Isotype control antibodies:
  • Rat IgG2a? (e.g., R35–95) is often used as a negative control to ensure specificity of MECA-367-mediated effects.
  • Mouse IgG3-FITC, IgG1-PE, IgG1-PerCP, and IgG1-APC are cited as isotype controls in multi-color flow cytometry panels that include MECA-367.
• PF-00547659:
  • A clinically developed anti-human MAdCAM-1 antibody. PF-00547659 is often compared or co-used with MECA-367 to study MAdCAM-related pathways, particularly for translational work between rodent and human systems.
• VCAM-1 (Vascular Cell Adhesion Molecule-1):
  • Included in the context of adhesion molecule studies and may be measured alongside MAdCAM-1 to assess differential roles in leukocyte trafficking and inflammation.
• Lymphocyte markers and ligands relevant to MAdCAM-1:
  • LPAM-1 (?4?7 integrin): The primary ligand for MAdCAM-1, often used in conjunction with MECA-367 to study lymphocyte migration and homing to gut-associated lymphoid tissues.
  • CD62L (L-selectin): Another molecule mediating lymphocyte homing that interacts with MAdCAM-1 and is frequently studied in similar experimental setups.

In summary, the most commonly used antibodies or proteins with MECA-367 in the literature are isotype controls (for specificity), related anti-MAdCAM-1 antibodies like PF-00547659, adhesion molecules like VCAM-1, and lymphocyte markers such as ?4?7 and CD62L when investigating lymphocyte trafficking in the context of mucosal immunity or inflammatory models.

Key findings from scientific literature citing clone MECA-367—a rat monoclonal antibody targeting murine MAdCAM-1 (mucosal addressin cell adhesion molecule-1)—can be summarized as follows:

• MECA-367 actively blocks MAdCAM-1, inhibiting lymphocyte homing to gut-associated lymphoid tissue. This blockade reduces the recruitment of ?4?7^+^ leukocytes to intestinal mucosa, an interaction critical in gastrointestinal inflammatory diseases.
• In preclinical models of colitis and other inflammatory states, MECA-367 administration led to reduced lymphocyte infiltration, decreased mucosal tissue destruction, and overall improvement in clinical signs of disease. These studies demonstrate that targeting the MAdCAM-1/?4?7 axis is a promising approach for treating inflammatory bowel disease (IBD).
• In vivo, MECA-367 binds mouse MAdCAM-1 with high affinity (K_d 5.1 pmol/L) and induces a dose-dependent increase in circulating ?7^+^ memory T-cells, with no change in ?7^- populations. This increase results from displacement of T-cells from mucosal tissue into the bloodstream—providing a biomarker for effective MAdCAM-1 blockade in both preclinical models (mice) and translational studies (nonhuman primates).
• Experimental work using radiolabeled MECA-367 confirmed its specificity for MAdCAM-1 on endothelial cells in vivo and enabled quantification of MAdCAM-1 expression in various immunological conditions.
• In an infection model, blockade of MAdCAM-1 with MECA-367 affected the outcome and mortality in immunized animals challenged with Pseudomonas aeruginosa. The intervention influenced survival curves and demonstrated that mucosal homing pathways can modulate not just inflammation but also immune defense mechanisms in the respiratory tract.

Overall, the citations of clone MECA-367 have played an instrumental role in:

• Elucidating the pathophysiological role of MAdCAM-1 in lymphocyte trafficking and gut immunity
• Validating antagonism of MAdCAM-1 as a therapeutic strategy in preclinical models of IBD and inflammation
• Facilitating the development and validation of translational biomarkers for the pharmacodynamic activity of MAdCAM-1 blocking agents in drug development.

No significant findings related to antibiotic resistance markers or mecA gene studies are linked to clone MECA-367, as those references pertain to unrelated molecular biology topics.