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

Clone H22 is used for two distinct purposes in in vivo mouse studies, depending on context: as a murine hepatoma cell line (H22) for creating liver cancer models, and as a monoclonal antibody (H22) targeting mouse IFN-? (Interferon gamma) for immunological research and modulation.

1. H22 Hepatoma Cell Line Use

• Purpose: The H22 cell line, derived from mouse liver tumors, is commonly injected into mice to create a robust liver cancer model for research on tumor biology, metastasis, drug efficacy, and immunotherapy.
• Typical Experimental Setup:
  • H22 cells are cultured, checked for viability, and injected (usually 1 million cells in 100 µL) subcutaneously or orthotopically into immunodeficient mice such as NOD/SCID or nude mice.
  • Tumor growth is monitored and mice are randomized into treatment cohorts to test anti-cancer drugs, investigate immune responses, or examine therapeutic interventions.
  • Studies can track tumor progression, examine immune cell dynamics (e.g., regulatory T cells, myeloid-derived suppressor cells in spleen, bone marrow, and tumor tissue), and assess the efficacy of chemotherapy agents or immunotherapies.

2. H22 Monoclonal Antibody Use

• Purpose: Clone H22 refers to an Armenian hamster monoclonal antibody specific for mouse IFN-?. It is used to neutralize or detect IFN-? in vivo, enabling the study of IFN-? function in mouse models of infection, autoimmunity, or cancer.
• Typical Applications:
  • The antibody can be administered to mice to block or deplete IFN-? activity, helping researchers understand the cytokine's role in immune responses or pathology.
  • It has been validated for in vivo use and is commonly used in functional studies requiring IFN-? modulation.
  • Researchers also use H22 antibody for flow cytometry or ELISA to detect and quantify IFN-? produced in mouse tissues.

Summary Table: H22 Uses in In Vivo Mouse Studies

If your question refers to one particular "H22" reagent, please specify whether you mean the cancer cell line or the IFN-? monoclonal antibody, as both are widely used and context-dependent.

Based on the search results, the correct storage temperature for sterile packaged clone H22 depends on the specific antibody and storage duration:

Short-term Storage

For short-term storage, clone H22 antibodies should be stored at 2°C to 8°C. This temperature range is suitable for up to one month from the date of receipt.

Long-term Storage

For long-term storage, clone H22 antibodies should be stored at -20°C to -70°C. This storage temperature maintains stability for up to 12 months from the date of receipt.

Important Storage Considerations

The storage guidelines emphasize several critical points for maintaining antibody integrity:

Avoid Freeze-Thaw Cycles: Use a manual defrost freezer and avoid repeated freeze-thaw cycles to prevent protein denaturation. For antibodies formulated with glycerol, storage at -20°C prevents freezing and eliminates freeze-thaw damage.

Sterile Handling: For functional grade preclinical antibodies that may be stored sterile, they can be kept at 2-8°C for up to one month as received. For longer storage periods, the antibodies should be aseptically aliquoted in working volumes without diluting and stored at ? -70°C.

Undiluted Storage: The antibody solution should be stored undiluted to maintain optimal stability.

The specific clone H22 mentioned in the search results includes both anti-mouse IFN-? antibodies and anti-human CD64 antibodies, but the storage requirements are consistent across both applications.

Other commonly used antibodies or proteins with H22 in the literature include monoclonal antibodies targeting CD64, such as M22, anti-TNF monoclonal antibodies (e.g., infliximab and adalimumab), and experimental proteins like single-chain antibody fragments (scFv) derived from H22 itself.

Supporting details:

• Anti-TNF monoclonal antibodies: H22 is frequently studied in combination with anti-TNF mAbs like infliximab and adalimumab to explore its capacity to block CD64 and prevent these antibodies from binding to activated immune cells. These experiments assess how H22 influences the capture and function of therapeutic antibodies.
• Other CD64-specific monoclonal antibodies: M22 is another full-length monoclonal antibody mentioned in the literature that binds to CD64. Both M22 and H22 recognize epitopes on CD64 independent of the Fc? domain, but are used for different experimental purposes.
• Experimental molecular fragments: H22-derived single-chain fragments (H22(scFv)) are often engineered and used for studies of CD64 blockade, gene delivery systems, and targeted therapies.
• Cell lines and detection reagents: Various detection antibodies (such as anti-His tag conjugates like anti-His5-Alexa Fluor 488 and goat anti-mouse PE) are used in flow cytometry to analyze H22 binding.
• Stimulatory cytokines: IFN-? (Interferon gamma) is commonly used to stimulate immune cell lines (such as HL-60) to increase CD64 surface expression and further evaluate H22’s effectiveness on activated cells.

Additional relevant proteins/antibodies used in H22 studies:

• Proteins involved in CD64 signaling or cell activation, including surface TNF (mTNF) and pro-inflammatory cytokines, are studied to determine H22’s downstream effects.
• Recombinant detection and purification tags (e.g., His10), as well as vectors like pET21a for molecular cloning and expression, are part of experimental protocols when working with H22-based constructs.

Summary Table:

Where the focus is disease or therapy research, anti-TNF monoclonal antibodies and other CD64-binding antibodies (such as M22) are primary co-reagents in studies involving H22.

The phrase "clone H22" appears in multiple scientific contexts, referring to different molecular and cellular entities. Here’s a synthesis of the key findings associated with "H22" in the literature, focusing on the relevant clone(s), and distinguishing between cell line, monoclonal antibody, and other usages.

H22 as a Hepatocellular Carcinoma Cell Line

Most scientific references to "H22" pertain to the hepatoma-22 (H22) cell line, a mouse model widely used in cancer research, particularly for hepatocellular carcinoma (HCC). Key findings from the H22 tumor-bearing mouse model include:

• Development of a Protocol: A detailed protocol was established using H22 tumor-bearing mice to screen potential anticancer targets, including the use of natural compounds (such as Ulva lactuca L. polysaccharide and 5-fluorouracil) and subsequent miRNA sequencing to identify novel therapeutic targets for HCC.
• Therapeutic Screening: The model allows for the evaluation of the anti-tumor effects of compounds through miRNA profiling, RT-qPCR, and Western blotting, aiding in the discovery of new miRNA-based therapies for liver cancer.
• Model Utility: H22 cells are injected into mice to establish tumors, which are then treated and analyzed, making this a robust platform for preclinical testing of HCC therapies.

H22 as a Monoclonal Antibody

A separate line of research identifies H22 as a monoclonal antibody (specifically, a recombinant single-chain variable fragment, H22(scFv)), with the following key findings:

• Target Specificity: H22(scFv) specifically binds to and blocks human CD64 (Fc?RI), a high-affinity IgG receptor, without activating it or inducing pro-inflammatory cytokine production.
• Functional Blockade: This antibody fragment prevents CD64 from capturing anti-TNF monoclonal antibodies (e.g., infliximab, adalimumab), thereby reducing unwanted immune activation that can occur with some therapeutic antibodies.
• Therapeutic Potential: Because H22(scFv) does not activate CD64 or trigger inflammation, it is considered a potential candidate for treating chronic inflammatory diseases where modulation of Fc receptor activity is desired.

H22 as an Anti-IFN? Antibody Clone

In immunology, clone H22 also refers to a monoclonal antibody against mouse interferon-gamma (IFN-?):

• Specificity: H22 recognizes an epitope on mouse IFN-?, a cytokine critical for innate and adaptive immunity against infections and tumors.
• Applications: This antibody is used in research for detecting and quantifying mouse IFN-?, commonly applied in techniques like Western blotting and functional assays.
• Biological Relevance: IFN-?, targeted by H22, is predominantly secreted by NK, NKT, and T cells and plays a pivotal role in antiviral and antitumor immunity.

DNA Vaccine Targeting H22 Tumor Cells

Another application involves constructing a DNA vaccine against domains 1–3 of flk-1 (VEGFR-2), which was shown to induce an immune response capable of blocking H22 tumor growth in mice by inhibiting angiogenesis, demonstrating the utility of H22 cells in immunotherapy research.

Summary Table

Conclusion

The key findings from "clone H22" citations depend on the context:

• As a tumor cell line, H22 is central to HCC research and therapeutic screening.
• As an antibody clone, H22 targets either CD64 (inhibiting Fc?RI function without activation) or IFN-? (for immunological assays).
• As a target for DNA vaccines, H22 tumors can be inhibited by immune strategies blocking angiogenesis.

Careful distinction is required when interpreting "H22" in scientific literature to avoid conflating these distinct biological entities.