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| Immunogen | Recombinant Protein |
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Product Overview
Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel Lindau protein (VLH) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7).HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8).
Validated Applications & Recommended Dilutions
| Application | Recommended Dilution |
|---|---|
| WB | 1:1000-2000 |
| IHC | 1:100-200 |
Antibody Specifications
| Target | HIF-1 β/ARNT |
|---|---|
| Host Species | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG |
| Conjugate | Unconjugated |
| Clone Number | 4C5 |
| Purification | Protein A Purification |
| Concentration | 1 mg/mL |
| Molecular Weight | Observed 87kDa |
Immunogen
Recombinant Protein UniProt: P27540.
Species Reactivity & Cross-Reactivity
Validated for: Mouse. Verify suitability in your sample type prior to use. For species or applications not listed in the datasheet, consult our technical team before purchase.
Storage & Handling
Store at -20℃ Valid for 12 months. Avoid freeze / thaw cycles.
Storage Buffer: PBS with 0.02% sodium azide, 0.5% protective protein and 50% glycerol, pH7.4
Related Products
Browse additional antibodies targeting HIF-1 β/ARNT and complementary reagents — including secondary antibodies, blocking buffers, and detection kits — available through BioHippo.
The recommended starting dilution for Western Blot is WB 1:1000-2000;IHC 1:100-200. Optimal dilution should be determined empirically for each laboratory setup, sample type, and detection system. Use freshly prepared lysates with protease inhibitors; add phosphatase inhibitors if studying phosphorylation-dependent forms of HIF-1 β/ARNT. If background is high, increase blocking (5% non-fat milk or BSA in TBST) and/or increase wash stringency.
For IHC-P, heat-induced epitope retrieval (HIER) is typically required. Recommended starting conditions: citrate buffer pH 6.0 (microwave or pressure cooker, ~20 min) for rodent samples; EDTA buffer pH 9.0 for human clinical samples. After retrieval, allow sections to cool gradually before proceeding to blocking. Include a HIF-1 β/ARNT-positive tissue control (confirmed by literature or database expression data) and a negative isotype control in every run. Recommended dilution: WB 1:1000-2000;IHC 1:100-200. Refer to the product datasheet for any target-specific retrieval guidance.
For intracellular targets including most nuclear and cytoplasmic proteins, fix cells with 4% paraformaldehyde (15 min, room temperature) followed by permeabilization with 0.1–0.2% Triton X-100 or saponin (5–10 min). For surface membrane targets, fixation without permeabilization may be sufficient. Block with 5–10% normal serum (matching the secondary antibody host species) for 30–60 min. Incubate primary antibody at the recommended dilution (WB 1:1000-2000;IHC 1:100-200) at 4°C overnight or 1–2 h at room temperature. Confirm nuclear/cytoplasmic localization of HIF-1 β/ARNT against published protein databases before designing your protocol.
This antibody has been experimentally validated in: Mouse. Cross-reactivity with other species has not been systematically evaluated. If your sample species is not listed, assess immunogen sequence homology to your target species ortholog. The immunogen used is: "Recombinant Protein" — higher sequence identity between species increases the probability of cross-reactivity, but experimental validation with appropriate controls is necessary before drawing scientific conclusions.
Store at -20℃ Valid for 12 months. Avoid freeze / thaw cycles. Storage Buffer: PBS with 0.02% sodium azide, 0.5% protective protein and 50% glycerol, pH7.4. To maintain antibody activity, avoid repeated freeze-thaw cycles. Aliquot into single-use volumes before first freeze. If working with the antibody frequently, keep a working stock at 4°C for up to 4 weeks — add 0.02% sodium azide to the working stock as a bacteriostatic agent if azide is not already present in the buffer. Monitor performance against a validated positive control when transitioning between storage aliquots or lots.
This monoclonal antibody (clone 4C5) provides highly consistent, epitope-specific recognition of HIF-1 β/ARNT, making it ideal for quantitative assays, multiplex panels, and studies requiring strict reproducibility across experiments and independent laboratories. Unlike polyclonal preparations, each production batch from a single hybridoma clone generates antibody with identical epitope specificity, affinity, and isotype — ensuring reliable signal comparison in longitudinal studies or multi-site collaborations. When citing this antibody in publications, include the clone designation (clone 4C5) to enable third-party reproducibility verification.
Customization & Add-ons: Can't find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.
