Anti-Mouse/Rat CD90.1 (Thy-1.1) [Clone OX-7] — Purified in vivo GOLD™ Functional Grade

Clone OX-7 is primarily used in in vivo mouse studies to target the CD90.1 (Thy1.1) alloantigen for cellular labeling, lineage tracking, and functional modulation of CD90.1-positive cells; it is most effective in mouse strains expressing CD90.1 (such as AKR and FVB), but not in those expressing CD90.2 (e.g., C57BL/6, BALB/c).

Key uses of OX-7 in in vivo mouse studies:

• Cell Depletion or Modulation: Administration of the OX-7 antibody can induce functional changes in CD90.1+ cells, such as activation or depletion, depending on the protocol and dose.
• Lineage Tracking/Marking: Using OX-7, researchers can distinguish CD90.1+ cells from CD90.2+ populations, facilitating adoptive transfer experiments or fate mapping in mixed-strain or transgenic settings.
• Functional Studies: OX-7 is used for investigating the role of Thy1.1+ cells in various immunological or neurological disease models, particularly where selective targeting of this epitope is required.

Additional notes:

• OX-7 binding can have biological effects—such as leukocyte activation, apoptosis of glomerular mesangial cells, induction of glomerular nephritis, and increased vascular permeability—which researchers should factor into study design.
• OX-7 does not react with CD90.2 (the predominant Thy1 isoform in most laboratory mouse strains, e.g., C57BL/6, BALB/c), which limits its application to CD90.1 strains or genetically modified cells expressing Thy1.1.
• In addition to immunological studies, OX-7 has been used for neural and stem cell tracking since CD90 is also expressed on mature neurons and some stem cell populations.

Summary Table: OX-7 use in in vivo mouse studies

All experimental designs using clone OX-7 must account for its specificity and possible functional/biological effects on target cells.

For the clone OX-7 antibody, the correct storage temperature is 2–8°C (refrigerated, not frozen), as specified for multiple commercially available versions of this monoclonal antibody (Pacific Blue-conjugated, APC-conjugated, and unconjugated formats).

• Do not freeze the antibody solution; freezing can damage the protein and reduce its efficacy.
• Protect from light during storage to prevent degradation.
• Aliquoting is recommended to avoid repeated freeze-thaw cycles, which can also degrade the antibody.

Always check the specific product datasheet for your lot, as recommendations can vary slightly depending on the formulation and supplier. However, for clone OX-7, refrigeration at 2–8°C is the standard and safest storage condition.

Commonly Used Companion Antibodies and Proteins with OX-7 (Anti-CD90)

OX-7 is a well-characterized monoclonal antibody that specifically targets rat CD90 (Thy-1), a glycosylphosphatidylinositol (GPI)-anchored, immunoglobulin superfamily member expressed on hematopoietic stem cells, neurons, thymocytes, and other cell types. In the literature and in laboratory practice, OX-7 is frequently used alongside other antibodies or proteins for co-staining, co-localization studies, and control experiments.

Common Companion Antibodies/Proteins

• Beta Tubulin (e.g., ab6046): Used as a loading control in immunocytochemistry (ICC) to provide a reference for cellular structure and normalization across samples. In a published protocol, OX-7 was used together with a rabbit polyclonal antibody to beta tubulin, with secondary antibodies labeled with different fluorophores (Alexa Fluor® 488 for mouse IgM and Alexa Fluor® 594 for rabbit IgG) to enable simultaneous visualization of CD90 and the cytoskeleton.
• Goat Anti-Mouse IgG (e.g., ab97040): Commonly employed as a secondary antibody in Western blotting to detect the primary OX-7 antibody.
• CD45: CD90 (Thy-1) has been shown to interact with CD45 in lymphocyte signaling, so antibodies against CD45 (a pan-leukocyte marker) are sometimes used in pathways or co-expression studies, although detailed protocol citations are less direct in the available sources.
• Pan-Cytokeratin (panCK), Cytokeratin (CK), CSVs: While these are not directly paired with OX-7 in the provided literature, they represent a set of widely used markers in immunofluorescence (particularly for circulating tumor cell (CTC) studies). Their strategic combination (e.g., panCK + CSV) is typical in multi-antibody panels for cell surface phenotyping, illustrating a common approach in the field even if not specifically with OX-7.
• General Isotype Controls: In flow cytometry and other immunoassays, isotype control antibodies (e.g., mouse IgG1, ?) are routinely used alongside OX-7 to assess non-specific binding.

Applications and Contexts

• Immunocytochemistry (ICC): OX-7 is paired with beta tubulin for dual labeling, allowing researchers to distinguish CD90 localization from the microtubule network.
• Western Blotting: OX-7, after primary incubation, is detected with anti-mouse IgG secondary antibodies.
• Flow Cytometry: In studies of rat splenocytes, OX-7 is used with appropriate secondary antibodies and isotype controls for specificity assessment.
• Multi-antibody Panels: The general practice in cell phenotyping is to use OX-7 within a larger panel, though specific companion antibodies depend on the biological question. For example, in hematopoiesis, markers like CD34, CD45, and lineage-specific antigens may be used alongside CD90.

Summary Table

Key Points

• Beta tubulin is a classic companion for OX-7 in ICC for structural reference and normalization.
• Secondary antibodies (anti-mouse IgG) are routinely used with OX-7 in Western blotting.
• Isotype controls and additional cell surface markers (e.g., CD45) are standard in flow cytometry and co-expression studies.
• While pan-cytokeratin and related markers are not specifically cited with OX-7, they exemplify the multi-marker panels common in cell phenotyping, a context in which OX-7 may also be used.

The specific choice of companion antibodies with OX-7 depends on the experimental design, but these examples reflect common practices in the literature.

Based on the scientific literature, clone OX-7 has generated several key findings across multiple research areas, demonstrating its utility as both a research tool and therapeutic intervention.

Neurological Research Findings

Clone OX-7 has shown significant promise in neuroscience research, particularly in promoting neural development. The antibody has been demonstrated to promote neurite outgrowths on peripherin-stained sympathetic rat neurons when examined using fluorescence microscopy. This finding suggests that CD90/Thy-1 plays an important role in neuronal development and potentially in nerve regeneration processes.

Nephrology and Disease Model Development

One of the most notable applications of clone OX-7 has been in creating experimental disease models. The antibody has been reported to induce glomerular nephritis in Wistar rats, making it a valuable tool for studying kidney disease mechanisms and testing potential therapeutic interventions. This finding has likely contributed to our understanding of immune-mediated kidney diseases and the role of CD90 in renal pathology.

Binding Affinity and Specificity Studies

Quantitative binding studies have established the high affinity of clone OX-7 for its targets. The Fab? fragment of MRC OX-7 demonstrates an affinity of 3 × 10? M?¹ for rat Thy1 and 3 × 10? M?¹ for mouse Thy1.1. These measurements provide crucial information for researchers regarding optimal concentrations and expected binding kinetics in experimental applications.

Cross-Species Reactivity Patterns

Clone OX-7 exhibits selective cross-reactivity with specific mouse strains, recognizing the mouse CD90.1 (Thy-1.1) alloantigen found in AKR/J and PL strains, but not CD90.2 (Thy-1.2) found in many other mouse strains. This specificity has been valuable for studying allelic differences and strain-specific responses in mouse models.

Broad Tissue Distribution Studies

Research has revealed that clone OX-7 recognizes CD90 across multiple tissue types and species. The antibody has been shown to cross-react with rabbit and guinea pig thymus, brain, and intestine, indicating the evolutionary conservation of CD90 epitopes and expanding its utility across different animal models.

Cellular Expression Profiling

Studies using clone OX-7 have mapped CD90 expression to diverse cell populations, including hematopoietic stem cells, early myeloid and erythroid cells, immature B lymphocytes, thymocytes, recent thymic emigrants, neurons, glomerular mesangial cells, endothelial cells at inflammatory sites, mast cells, and dendritic cells. This comprehensive expression profile has enhanced our understanding of CD90's role in various physiological processes.

These findings collectively demonstrate that clone OX-7 has been instrumental in advancing research across neuroscience, immunology, nephrology, and stem cell biology, while also serving as a reliable tool for flow cytometry, immunohistochemistry, and other analytical techniques.