Anti-Mouse LPAM-1 – Purified in vivo GOLD™ Functional Grade

Clone DATK32 is widely used in mice for in vivo applications that primarily focus on the blockade of α4β7 integrin (LPAM-1)-mediated lymphocyte adhesion and trafficking, especially in models of mucosal immunity and inflammation.

Key in vivo applications include:

• Blocking lymphocyte homing to the gut and associated lymphoid tissues: By inhibiting the interaction between α4β7 (the target of DATK32) and its ligands (notably MAdCAM-1 on gut endothelial cells), DATK32 prevents lymphocytes from migrating into the intestinal mucosa—this is a central mechanism in studies of gut-specific immune responses and inflammatory bowel disease (IBD).
• In vivo functional studies of lymphocyte trafficking: DATK32 is used to dissect the role of α4β7 integrin in trafficking under various inflammatory or homeostatic conditions by selectively blocking this pathway and observing immune cell recruitment and localization.
• Preclinical models of IBD and mucosal inflammation: DATK32 has been used in mouse models (such as dextran sulfate sodium (DSS)-induced colitis) to examine the therapeutic potential and the mechanistic importance of α4β7 integrin in disease progression, often paralleling the action of vedolizumab (a clinically used anti-α4β7 mAb).

Other documented in vivo uses:

• Radioimmune imaging: Labeled DATK32 can be used for noninvasive imaging of α4β7+ lymphocyte recruitment to inflamed tissues, aiding in the assessment of disease activity in models such as DSS-induced colitis.
• Immunohistochemistry and ex vivo tissue analysis: After in vivo administration, subsequent tissue collection and staining can be used to validate leukocyte distribution and correlate with histopathological findings.

DATK32 is considered the standard tool antibody for in vivo blockade experiments targeting lymphocyte migration to gut-associated tissues in the mouse, and is also employed for mechanistic studies exploring the function of LPAM-1 (α4β7 integrin) in lymphocyte-endothelial interactions during inflammation.

In summary, clone DATK32 is most commonly used in vivo for:

• Functional blockade of α4β7-dependent cell adhesion and lymphocyte trafficking
• Preclinical studies of intestinal inflammation and immunity
• Imaging and mechanistic analysis of gut homing in mice

These applications are central to understanding mucosal immunity and developing therapeutics for related inflammatory diseases.

The most commonly used antibodies or proteins with DATK32 (anti-mouse LPAM-1, integrin α4β7) in the literature include:

• Natural ligands:

  • VCAM-1 (CD106)
  • MAdCAM-1
  • Fibronectin
    These are frequently used in cell adhesion assays to study binding or blocking activity of α4β7.

• Other integrin subunit antibodies:

  • CD49d (Integrin α4)
  • CD103 (Integrin αE)
    Antibodies against related integrins (like αEβ7/HML-1) help to define lymphocyte subsets and distinguish trafficking pathways.

• Lineage or marker antibodies:

  • CD45R (B220) for B cells
  • CD3, CD4, CD8 for T cell subsets
    Frequently included in multi-color flow cytometry panels to phenotypically characterize lymphocyte populations alongside DATK32.

• Isotype controls:Used as negative controls for flow cytometry or functional assays to confirm specificity.

• Therapeutic analogs:

  • Vedolizumab (humanized anti-α4β7) and related agents
    Sometimes used for comparative studies of blocking or functional effects.

Additional commonly referenced proteins/antibodies:

• FIB504: Another antibody clone targeting β7 integrin, sometimes compared or used with DATK32.
• E-cadherin (CD324): Used in studies involving αEβ7 integrin and its ligand, especially in mucosal immunity research.
• TNFα: Occasionally included in inflammation-related studies assessing α4β7+ lymphocyte trafficking.

These combinations facilitate studies of lymphocyte adhesion, homing, and mucosal immune responses, with DATK32 being central to the detection or functional blocking of α4β7 integrin in mouse models.

Clone DATK32 is a rat monoclonal antibody widely cited in scientific literature as a selective blocker and detector of mouse integrin α4β7 (LPAM-1), an adhesion molecule essential for lymphocyte trafficking, particularly to mucosal tissues.

Key findings and uses reported in the literature include:

• Detection and Quantification: DATK32 is validated for flow cytometry, immunohistochemistry, and immunoprecipitation to detect LPAM-1 on mouse lymphocytes, helping researchers characterize immune cell populations, especially those trafficking to gut-associated lymphoid tissue.

• Blocking/Neutralization: DATK32 reliably blocks α4β7 integrin-mediated adhesion to its natural ligands, including VCAM-1, MAdCAM-1 (which is gut-expressed), and fibronectin, both in vitro and in vivo. This blockade is foundational in studies of lymphocyte homing and inflammatory responses, especially in gut immunity and inflammation.

• Functional Effects: The antibody has been shown to induce aggregation of certain lymphocyte lines (such as the mouse CD8+ T cell lymphoma TK1) and block their adhesion, confirming its activity in modulating integrin function.

• Research Applications:

  • Gut Immunology: DATK32 blockade is used to study lymphocyte homing to Peyer’s patches and other gut lymphoid tissues, modeling mechanisms relevant for therapies like vedolizumab, an anti-α4β7 integrin antibody for IBD.
  • Atherosclerosis: Elevated α4β7 expression detected by DATK32 has been observed at murine atherosclerotic lesions, implicating LPAM-1 in vascular inflammation beyond traditional gut contexts.
  • Imaging and Diagnostics: Radio-labeled DATK32 is utilized for imaging α4β7 expression in vivo, demonstrating stability and specificity, and opening diagnostic applications, particularly in inflammation and colitis models.

• Impact on Experimental Design: The clear specificity and robust blocking ability of DATK32 have made it the gold standard for dissecting the role of α4β7 in experimental mouse models of immune cell trafficking, inflammation, and related pathologies.

In summary, clone DATK32 is essential for mechanistic studies of mouse α4β7 integrin, providing both a tool for immune cell identification and a means to interrogate the function of lymphocyte trafficking—especially in models of gut inflammation, mucosal immunity, and, more recently, vascular disease.

Dosing regimens for clone DATK32 (anti-mouse α4β7 integrin antibody) differ based on route of administration, disease model, and experimental goals across mouse studies. The most commonly reported regimens include:

• Intraperitoneal (IP) injection: 25 mg/kg every 3 days (Q3D) in DSS acute colitis models.
• Intracolic (IC) dosing: 25 mg/kg every 3 days (Q3D) or 25 mg/kg every day (QD); a lower dose, 5 mg/kg, has been administered IC daily.
• IP in EAE (experimental autoimmune encephalomyelitis) model: 10 mg/kg twice weekly in C57BL/6 mice for CD4⁺ T cell depletion.

Supporting details:

• In DSS-induced acute colitis:

  • IP DATK32 at 25 mg/kg Q3D and IC DATK32 at 25 mg/kg Q3D or QD were compared, as well as a lower IC dose (5 mg/kg QD). Drug concentrations in colon tissue and content were higher for IC compared to IP, especially with more frequent administration.
  • A volume of 0.1mL/20g mouse was used for dosing from day 0 to day 14.

• In EAE models (autoimmunity, C57BL/6 mice):

  • DATK32 was administered IP at 10 mg/kg twice weekly to deplete CD4⁺ T cells.

Variation by mouse model and dose:

• Colitis/IBD models (often use higher doses, IP or IC, with dosing frequency based on inflammation timeline).
• Autoimmunity models (EAE) (use lower dose, IP, tailored to T cell depletion needs).

Route & frequency effects:

• IC administration yields higher local drug concentrations in colon tissue versus IP, and frequent dosing (QD vs Q3D) further increases these levels.
• IP administration is standard for systemic effects, as seen in EAE and other depletion studies.

Additional considerations:

• Actual regimens may be further optimized based on specific experimental outcomes, mouse strain, or disease pathology, with investigators advised to determine optimal dosing empirically for their application.

Summary Table:

For other disease models or strains, dosing may require further individual optimization depending on research aims and pharmacokinetic parameters.