Anti-Mouse IL-1α [Clone ALF-161] — Purified in vivo GOLD™ Functional Grade

Clone ALF-161 is primarily used in in vivo mouse studies to neutralize the activity of interleukin-1 alpha (IL-1?), a key pro-inflammatory cytokine implicated in immune and inflammatory responses.

In practice, ALF-161 is administered to mice (often via intravenous or intraperitoneal injection) prior to or during experimental procedures where IL-1?'s role is being investigated. For example:

• Studies of Sterile Inflammation: Mice receive ALF-161 antibody (e.g., 200 ?g intravenously) to specifically block IL-1?, allowing researchers to examine its contribution to inflammation after a sterile injury, such as acetaminophen-induced liver injury.
• Viral or Immune Challenges: ALF-161 is injected (alongside other antibodies) to assess IL-1?'s requirement in T cell-driven phenotypes like weight loss during respiratory viral infection.
• General Neutralization Studies: The clone is validated for functional blockade in various in vivo models, enabling researchers to define the physiological and pathophysiological roles of IL-1?-neutrophil, macrophage, and lymphocyte interactions.

Key technical details:

• ALF-161 is an Armenian hamster IgG1 monoclonal antibody designed for high specificity and low endotoxin content (making it suitable for in vivo work).
• It recognizes precursor, secreted, and membrane-associated forms of mouse IL-1?.
• The antibody can neutralize natural or recombinant IL-1?, thus blocking IL-1?-mediated effects in experimental mice.
• Typical protocols recommend i.v. or i.p. injection at doses around 200 ?g per mouse, with timing adjusted to study design (e.g., 1 hour before injury/antigen challenge).

Applications include:

• Functional blockade in in vivo inflammation models
• Use in cytokine neutralization studies
• Supporting experimentation in autoimmunity, infection, or injury models where IL-1?'s function must be isolated.

In summary, clone ALF-161 enables researchers to explore the role of IL-1? by selectively neutralizing its activity in living mice, proving essential for mechanistic studies of inflammation and immune regulation.

The correct storage temperature for sterile packaged clone ALF-161 depends on both the product form and the intended storage duration:

• Short-term storage (up to 1 month): Store at 2°C to 8°C (refrigerator temperature).
• Long-term storage: For periods longer than one month, aliquot and store at -20°C (or lower, such as ? -70°C, depending on manufacturer instructions). Avoid repeated freeze-thaw cycles to prevent antibody degradation.

For fluorophore-conjugated forms (e.g., PE- or FITC-conjugated ALF-161), specifically do not freeze; keep these at 2°C to 8°C and protect from light to prevent loss of function.

In summary:

• Sterile, unconjugated antibody: 2–8°C (short term), -20°C or below (long term, aliquoted, avoid freeze-thaw).
• Fluorophore-conjugated antibody: 2–8°C only, do not freeze.

Check your product label or Certificate of Analysis for any manufacturer-specific recommendations.

The antibody ALF-161 is most commonly used for detection of mouse interleukin-1 alpha (IL-1?) in a range of immunological assays. In the literature and standard assay protocols, ALF-161 is almost always paired with other antibodies or proteins specific to IL-1?, as well as with assay controls and standards. The most frequently used additional antibodies or proteins include:

• Biotinylated Poly5034 antibody: Used as the detecting antibody in sandwich ELISA or ELISPOT assays, with ALF-161 serving as the capture antibody.
• Biotin anti-mouse IL-1? polyclonal antibody (13-7111): Utilized as the detection antibody in ELISAs where ALF-161 is the capture antibody.
• Recombinant mouse IL-1? protein (14-8011): Frequently used as a standard or blocking antigen for validating staining specificity or quantifying assay results.

In flow cytometry and immunofluorescence applications involving ALF-161:

• Isotype controls (e.g., Armenian hamster IgG isotype controls like PE-G235-2356): Used to establish background staining levels.
• Blocking antibody controls (unlabeled ALF-161): Used to confirm specificity of staining by pre-blocking with the same antibody or excess recombinant IL-1?.

For neutralization, ALF-161 may be used alone to block IL-1? bioactivity, but studies often also measure related cytokines such as:

• IL-1?: Since IL-1? and IL-1? share functional pathways but are distinct proteins with different antibodies.

In summary, common pairings in the literature and commercial protocols with ALF-161 include:

• Capture-detection pairs: ALF-161 (capture) + biotin Poly5034 or 13-7111 (detection).
• Standards and controls: Recombinant IL-1? protein, Armenian hamster IgG isotype control, unlabeled ALF-161.
• Related cytokine antibodies: Often, parallel measurement of IL-1? using its specific antibodies is performed for comprehensive cytokine profiling.

No evidence was found for routine co-use with unrelated proteins or markers, unless included for multiparameter flow cytometry where additional surface or intracellular markers may be used for cellular phenotyping (not IL-1?-specific).

Clone ALF-161 is a widely used monoclonal antibody against mouse interleukin-1 alpha (IL-1?), and it has been cited in multiple scientific studies for its roles in immunology and cancer research. Key findings from the scientific literature that cite ALF-161 include:

• Neutralization and Detection of IL-1?: ALF-161 can neutralize natural or recombinant IL-1?, and is also used as a capture antibody in ELISA and ELISPOT assays for detecting IL-1? in mouse biological samples. This establishes its utility in both functional studies and cytokine quantification.

• Role of IL-1? in Tumor Immunity and Immunotherapy Resistance:

  • IL-1? drives resistance to immunotherapy in murine melanoma. In preclinical mouse models, neutralization of IL-1? (with antibodies such as ALF-161)—but not IL-1?—prolongs survival and enhances the efficacy of various immunotherapies (e.g., anti–PD-L1, peptide vaccination, ISF35).
  • Blockade of IL-1 signaling, including specifically IL-1?, increases the number of tumor-specific CD8+ T cells and IFN-?–producing T cells within the tumor microenvironment, suggesting that IL-1? acts as an immunosuppressive factor limiting effective anti-tumor immunity.

• Distinct Roles of IL-1? and IL-1?: Genetic and pharmacological blockade experiments show that only IL-1?, not IL-1?, mediates resistance to immunotherapy in the B16 melanoma mouse model, highlighting a non-redundant role for IL-1? in tumor-induced immune suppression.

• Hematopoietic and Myeloid Cell Function: IL-1? (detected/neutralized with ALF-161) mediates a broad range of immune and inflammatory responses, including myeloid cell activation and aging-related alterations in hematopoiesis that can promote tumor growth. Age-related enhancement of myelopoiesis via IL-1R1 signaling (stimulated by IL-1?) is implicated as a driver of cancer progression in aging mouse models.

• Wider Applications: The ALF-161 clone is validated for several applications—including in vivo neutralization, ELISA, ELISPOT, neutralization, and Western blotting—providing an essential tool for mechanistic immunology studies in mice.

Summary table of ALF-161–related findings:

In summary, clone ALF-161 is critical in demonstrating the immunosuppressive role of IL-1? in the tumor microenvironment, especially regarding resistance to immune checkpoint therapies in mice, and remains a key reagent for quantitative and functional studies of IL-1? in mouse immunology and cancer research.