Anti-Mouse IFNγ [Clone H22] — Purified in vivo PLATINUM™ Functional Grade

Clone H22 is primarily used in in vivo mouse studies as a neutralizing monoclonal antibody against mouse interferon-gamma (IFN-?) to block or inhibit IFN-? activity. This allows researchers to study the roles of IFN-? in various immune responses and disease processes by selectively suppressing its signaling.

Key applications and details:

• H22 (Hamster anti-Mouse, IgG1 ?) specifically binds to mouse IFN-? and not to other interferons like IFN-? or IFN-?, nor to human IFN-?.
• It inhibits IFN-?-mediated activities in vivo, such as antiviral responses, induction of class II MHC antigen, and macrophage-activating factor (MAF) functions.
• H22 neutralizes IFN-? in living animals, helping determine the cytokine’s contributions to immunity, inflammation, and disease models—especially in immunology and cancer studies.
• In mouse model experiments, H22 has been shown to block the binding of recombinant IFN-? to IFN-? receptor-positive cells, effectively reducing downstream IFN-? signaling and allowing observation of the effects of IFN-? absence.

In summary:

• Clone H22 is injected into mice to neutralize endogenous IFN-?, providing a mechanistic tool to dissect the role of this key cytokine in vivo.
• This approach is commonly used in studies on autoimmune diseases, tumor immunity, infectious diseases, and transplantation, wherever modulation of IFN-? responses is relevant to the experimental question.

For the sterile packaged clone H22, which is associated with different targets (e.g., mouse IFN-? and human CD64), the storage temperature can vary depending on the specific application and packaging. However, general guidelines for monoclonal antibodies apply:

• Short-term storage (up to a few weeks): Store at 2°C to 8°C. This is suitable for maintaining the antibody's efficacy without extensive degradation.

• Long-term storage: For extended periods, it is recommended to store the antibodies at temperatures ranging from -20°C to -70°C or even -80°C to maximize stability and prevent microbial growth.

Avoid repeated freeze-thaw cycles, as they can denature proteins and decrease antibody activity. It is important to follow the specific storage instructions provided with the product, as these may vary based on formulation and intended use.

Commonly used antibodies or proteins in the literature with H22 depend on which H22 you are referring to, as there are two widely used antibodies with this designation:

• Anti-human CD64 (Fc?RI) antibody: H22 and its derivatives (e.g., H22(scFv)), which target human CD64.
• Anti-mouse IFN-? antibody: Clone H22, which binds to mouse interferon gamma.

Below are key reagents frequently used in combination with or in the context of these two H22 antibodies:

For anti-human CD64 (H22 or H22(scFv)) studies:

• Anti-TNF monoclonal antibodies such as infliximab and adalimumab
  Used together with H22(scFv) to study the blockade of CD64-mediated anti-TNF capture in various cell-based assays.

• Detection antibodies
  Secondary reagents such as anti-His tag antibodies (e.g., anti-His5-Alexa Fluor 488) or goat anti-mouse antibodies (GAM-PE) are used to detect His-tagged H22(scFv) via flow cytometry.

• Cell surface and activation markers
  Other antibodies against cell surface proteins, such as those detecting HL-60 cell markers or induction with IFN-?, are used to stimulate or analyze myeloid cell responses.

• Cytokine detection antibodies
  To evaluate cytokine expression (including TNF), commercial anti-TNF antibodies and ELISA kits are sometimes paired with H22 in functional assays.

• Fusion proteins or conjugates
  CD64-targeted delivery tools: H22(scFv) has been fused to protein toxins or to gene delivery vehicles such as oligo-nona-arginine (9R) for targeted therapy or siRNA delivery in experimental oncology and immunology.

For anti-mouse IFN-? (clone H22) studies:

• Cytokine detection cocktails
  The H22 clone is commonly used alongside anti-IL-2, anti-TNF, and anti-IL-4 in multiparametric ELISA, intracellular cytokine staining, and flow cytometry to measure mouse immune responses.

• Secondary detection antibodies
  Anti-mouse IgG or anti-hamster secondary antibodies may be used for Western blotting, ELISA, or immunoprecipitation assays, in which H22 is used as the primary antibody.

• Co-stimulatory or cell phenotyping markers
  In flow cytometric analysis of T cell activation, H22 (anti-IFN-?) antibodies can be used with antibodies to surface markers such as CD4, CD8, CD3, or activation markers (CD44, CD25) for immune profiling.

Summary Table

The specific choice of reagents depends on the experimental context—whether the goal is blocking, detection, delivery, or immune profiling. The most frequently paired antibodies are anti-TNF for CD64 H22 studies, and anti-IL-2 or anti-TNF for mouse IFN-? H22 studies.

Key findings from scientific literature regarding clone H22 primarily refer to two unrelated biological contexts: the murine H22 hepatoma cell line (used in liver cancer studies in mice) and the E. coli ST131-H22 clone (a notable pathogenic lineage in microbiology). The most frequent citations concern the murine H22 hepatoma model. Below are the major findings from the literature as represented in your search results:

Murine Ascites Hepatoma Clone H22:

• Metastatic Potential and nm23 Expression:
  The study of H22 clones with varying lymphatic metastatic potential revealed that levels of nm23 mRNA correlate with metastatic behavior in these hepatomas. However, significant heterogeneity exists, suggesting that nm23 expression may be associated with—but not solely determinant of—metastatic capability in H22-derived cancers.

• Immune Suppression in Tumor Progression:
  In H22 tumor-bearing mice, the spleen is dominated by myeloid-derived suppressor cells (MDSCs) rather than regulatory T cells (Tregs) during tumor progression. MDSCs negatively correlate with T and NKT cells in the spleen and can significantly suppress immune reactivity (demonstrated by reduction of IFN-? in co-culture assays), indicating a principal role for MDSCs in immune evasion by H22 tumors.

• Target Discovery for Hepatocellular Carcinoma (HCC):
  The H22 model is valuable for screening therapeutic targets for HCC. Protocols detail establishing H22 tumor-bearing mice and applying treatments (such as Ulva lactuca L. polysaccharide or 5-fluorouracil), followed by analysis of changes in miRNA profiles. The model enables the identification and validation of potential miRNA targets relevant to cancer therapy.

E. coli ST131-H22 Clone:

• Significance as a Pathogen:
  The ST131-H22 clone is a major subgroup of E. coli ST131 responsible for extraintestinal infections (mainly urinary tract infections). Genomic analysis shows clinical isolates are closely related to animal (meat) sources, often carrying unique avian-associated ColV plasmids. Findings support the concept that a significant subset of community-acquired UTI cases in the US may be foodborne, with reservoirs in poultry dating back decades.

Summary Table: H22 Clone Findings

The majority of H22 citations in biomedical research relate to mouse liver tumor models and their mechanisms of metastasis and immune interaction, while citations in microbiology refer to the E. coli H22 lineage as an important foodborne pathogen. Always consult the specific subject context when encountering clone H22 in scientific literature.