Caspase-1, also known as IL-1 beta -converting enzyme (ICE), is an aspartic protease that plays a key role in the inflammatory response and apoptosis. Caspase-1 precursor (about 50kDa) can be cleaved and the active enzyme consists of a complex of two 20 kDa (aa 120-297) and two 10 kDa (aa 317-404) subunits which associate following cleavage of inactive precursors. Caspase-1 is required for proteolytic cleavage of the IL-1 beta precursor to form the active proinflammatory cytokine. Alternate splicing generates several additional Caspase-1 isoforms with deletions in the propeptide regions or also in the mature subunits. Within the large subunit, human Caspase 1 shares 61% aa sequence identity with mouse and rat Caspase-1.
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human
Cited:
Human, Rat
Applications
Validated:
Knockout Validated, Western Blot, Immunocytochemistry, Simple Western
Cited:
Immunohistochemistry, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human Caspase-1
Asn120-Asp297
Accession # P29466
Asn120-Asp297
Accession # P29466
Specificity
Detects human Caspase-1 in Western blots.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human Caspase‑1 Antibody
Detection of Human Caspase‑1 by Western Blot.
Western blot shows lysates of A431 human epithelial carcinoma cell line and THP-1 human acute monocytic leukemia cell line. PVDF Membrane was probed with 0.25 µg/mL of Goat Anti-Human Caspase-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6215) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Caspase-1 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Caspase‑1 in THP‑1 Human Cell Line.
Caspase-1 was detected in immersion fixed THP-1 human acute monocytic leukemia cell line using Goat Anti-Human Caspase-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6215) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human Caspase‑1 by Simple WesternTM.
Simple Western lane view shows lysates of A431 human epithelial carcinoma parental cell line and Caspase-1 knockout A431 cell line (KO), loaded at 0.2 mg/mL. A specific band was detected for Caspase-1 at approximately 52 kDa (as indicated) using 25 µg/mL of Goat Anti-Human Caspase-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6215) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human Caspase‑1 Specificity by Using Knockout Cell Line.
Western blot shows lysates of A431 human epithelial carcinoma parental cell line and Caspase-1 knockout A431 cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Caspase-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6215) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Caspase-1 at approximately 50 kDa (as indicated) in the parental A431 cell line, but is not detectable in knockout A431 cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Applications for Human Caspase‑1 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line
Knockout Validated
Caspase‑1
is specifically detected in A431 human epithelial carcinoma parental cell line but is not detectable in
Caspase‑1 knockout A431 cell line.
Simple Western
25 µg/mL
Sample: A431 human epithelial carcinoma cell line
Sample: A431 human epithelial carcinoma cell line
Western Blot
0.25 µg/mL
Sample: A431 human epithelial carcinoma cell line and THP‑1 human acute monocytic leukemia cell line
Sample: A431 human epithelial carcinoma cell line and THP‑1 human acute monocytic leukemia cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Caspase-1
Additional Caspase-1 Products
Product Documents for Human Caspase‑1 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Caspase‑1 Antibody
For research use only
Related Research Areas
Citations for Human Caspase‑1 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
NOD-like Receptor Signaling Pathways
