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-1α), 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-1α's role is being investigated. For example:

• Studies of Sterile Inflammation: Mice receive ALF-161 antibody (e.g., 200 µg intravenously) to specifically block IL-1α, 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-1α'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-1α–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-1α.
• The antibody can neutralize natural or recombinant IL-1α, thus blocking IL-1α–mediated effects in experimental mice.
• Typical protocols recommend i.v. or i.p. injection at doses around 200 µg 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-1α's function must be isolated.

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

The antibody ALF-161 is most commonly used for detection of mouse interleukin-1 alpha (IL-1α) 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-1α, 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-1α polyclonal antibody (13-7111): Utilized as the detection antibody in ELISAs where ALF-161 is the capture antibody.
• Recombinant mouse IL-1α 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-1α.

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

• IL-1β: Since IL-1α and IL-1β 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-1α protein, Armenian hamster IgG isotype control, unlabeled ALF-161.
• Related cytokine antibodies: Often, parallel measurement of IL-1β 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-1α-specific).

Clone ALF-161 is a widely used monoclonal antibody against mouse interleukin-1 alpha (IL-1α), 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-1α: ALF-161 can neutralize natural or recombinant IL-1α, and is also used as a capture antibody in ELISA and ELISPOT assays for detecting IL-1α in mouse biological samples. This establishes its utility in both functional studies and cytokine quantification.

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

  • IL-1α drives resistance to immunotherapy in murine melanoma. In preclinical mouse models, neutralization of IL-1α (with antibodies such as ALF-161)—but not IL-1β—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-1α, increases the number of tumor-specific CD8+ T cells and IFN-γ–producing T cells within the tumor microenvironment, suggesting that IL-1α acts as an immunosuppressive factor limiting effective anti-tumor immunity.

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

• Hematopoietic and Myeloid Cell Function: IL-1α (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-1α) 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-1α 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-1α in mouse immunology and cancer research.

Published dosing regimens for clone ALF-161 in mouse models are not extensively detailed in available sources, and reported data primarily reference its use in vitro or as a capture antibody for analytical applications. However, some in vivo research on mouse models, including studies of IL-1α blockade, cite ALF-161 as a key reagent for functional assays.

Key points from available information:

• In vitro neutralization: The ALF-161 antibody inhibits 50% of the biological effects of 4.0 ng/mL mouse IL-1α at a concentration of 0.031 µg/mL in a D10 cell proliferation assay.
• ELISA protocols: Typical concentrations used for sandwich ELISA capture range from 1–4 µg/mL.
• Reported applications: ALF-161 is validated for capture, ELISPOT, neutralization (functional grade), and intracellular staining for flow cytometry, but specific in vivo dosing (µg or mg per mouse per administration) is not reported in these sources.

In vivo usage:

• While the antibody is referenced in the context of mouse cancer models for IL-1α pathway disruption, neither the dose, frequency, nor route of administration (e.g. intraperitoneal, intravenous) is specified in the accessible details.
• Comparable antibody dosing regimens (for related murine cytokine studies, not specifically ALF-161) often use 100–500 µg/mouse i.p. between 1–3 times per week depending on the experimental aim and target. This suggests that researchers may extrapolate from such standards when employing functional neutralizing antibodies for in vivo intervention.

Variability by mouse model:

• No specific data compares dosing regimens of ALF-161 across different mouse strains or disease models (e.g., C57BL/6 versus BALB/c, tumor versus inflammatory disease).
• In related experiments with genetic knockout models, researchers unify genetic backgrounds to reduce dosing discrepancies, implying that, when specifics are unavailable, dose-finding studies (starting in the 100–500 µg/mouse range) are typically required for each model.

Summary table: Existing data on ALF-161 dosing/applications

Conclusion:
There is no published standardized dosing regimen for clone ALF-161 in various mouse models. Most guidance must be extrapolated from in vitro protocols or general murine antibody dosing standards, typically starting in the 100–500 µg/mouse i.p. range for functional blockade studies, with actual dosing requiring pilot calibration for each mouse strain and disease context. If specific experimental details are needed (model, disease, endpoint), researchers are advised to consult the manufacturer or recent publications for dosing optimization in their chosen system.