Caspase-1 Antibody (14F468) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-33230

Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 14F468

Format

Azide and BSA Free
View all Caspase-1 Primary Antibodies »

Product Specifications

Immunogen

Caspase-1 Antibody (14F468) - Azide and BSA Free was developed against a synthetic peptide corresponding to amino acids 371-390 RKVRFSFEQPDGRAQMPTTE of human caspase-1.

Reactivity Notes

Immunogen's sequence similarity with other species: Porcine (85%), Equine (80%), Canine (70%). Rat reactivity reported in scientific literature (PMID: 22133203).

Localization

Cytoplasm, Nucleus

Specificity

Caspase-1 Antibody (14F468) will recognize full-length Caspase-1 and cleaved caspase-1 forms that retain amino acids 371-390 of the Caspase-1 protein.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

45.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Caspase-1 Antibody (14F468) - Azide and BSA Free

Western Blot Analysis of Caspase-1 in Infected Mouse BMMs

Western Blot Analysis of Caspase-1 in Infected Mouse BMMs

P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in murine macrophages. BMM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and the activation of inflammasome components in the lysates [or supernatants (sup) where indicated] measured after 24 h by Western blot; beta-actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. Image collected and cropped by CiteAb from the following publication (https://journal.frontiersin.org/article/10.3389/fcimb.2017.00351/full), licensed under a CC-BY license. Image using the standard format of this product.
Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Intestine

Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Intestine

Tissue section of human intestine using Caspase-1 antibody (clone 14F468) at 5ug/ml concentration (1:200 dilution). The primary antibody binding to Caspase 1 in cells was detected using HRP conjugated anti-Mouse secondary antibody with DAB reagent, and the sections were further counterstained with hematoxylin for labeling cellular nuclei. This Caspase 1 antibody generated a diffused but specific cytoplasmic staining in columnar epithelia cells of villi, and a few cells depicted nuclear staining also. Only a subset of connective tissue cells in lamina propria depicted positivity (cytoplasmic) for this protein. Image using the standard format of this product.
Western Blot Detection of Caspase-1 in Human and Mouse Cell Lysates

Western Blot Detection of Caspase-1 in Human and Mouse Cell Lysates

Analysis in human HeLa (A) and mouse NIH3T3 lysate probed with Caspase-1 antibody at 0.5 ug/ml and 2 ug/ml, respectively.
Detection of Caspase-1 in LukAB Intoxicated THP1 Cells by Western Blot

Detection of Caspase-1 in LukAB Intoxicated THP1 Cells by Western Blot

LukAB is a potent activator of Caspase 1. THP1 cells were intoxicated with 50 ng/mL LukAB for 1 hour and cell lysates were analyzed by immunoblot for Caspase 1 cleavage, which indicates activation. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.ppat.1004970) licensed under a CC-BY license. Image using the standard format of this product.
Western Blot Analysis of Caspase-1 in Infected Human MDMs

Western Blot Analysis of Caspase-1 in Infected Human MDMs

P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in human macrophages. MDM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and the activation of inflammasome components in the lysates was measured after 24 h by Western blot; beta-actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. Image collected and cropped by CiteAb from the following publication (https://journal.frontiersin.org/article/10.3389/fcimb.2017.00351/full), licensed under a CC-BY license. Image using the standard format of this product.
Immunohistological Staining of Caspase-1 in Paraffin Embedded Adenocarcinoma of the Rectum

Immunohistological Staining of Caspase-1 in Paraffin Embedded Adenocarcinoma of the Rectum

Analysis in adenocarcinoma of the rectum (5 ug/ml), peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.
Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Colon

Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Colon

Analysis in a section of normal human colon using 5 ug/ml concentration. Distinct cytoplasmic staining along with some nuclear positivity was observed in crypts/mucosa, and staining was found to be more intense in the absorptive columnar epithelial cells. [10X Magnification]
Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Lung

Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Lung

Analysis in a section of normal lung from human using 5 ug/ml concentration. In this representative lung section, different type of cells including pseudostratified columnar epithelium of bronchiole and the simple squamous epithelium of alveoli may be seen to develop immunoreactivity for Caspase 1. [10X Magnification]
Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Skin

Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Skin

Analysis in a section of normal skin from human using 5 ug/ml concentration. Strong cytoplasmic/nuclear staining developed in all the epidermal cells, blood vessels and some cells of the dermal connective tissues layer. [10X Magnification]
Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Ovarian Cancer

Immunohistological Staining of Caspase-1 in Paraffin Embedded Human Ovarian Cancer

Analysis in a section of human ovarian cancer using 5 ug/ml concentration.
Detection of Caspase-1 in HeLa Cell Lysate by Simple Western

Detection of Caspase-1 in HeLa Cell Lysate by Simple Western

Analysis in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for Caspase-1 Antibody (14F468) - Azide and BSA Free

Application
Recommended Usage

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Paraffin

1:10-1:500

Simple Western

1:50

Western Blot

0.5-2 ug/ml
Application Notes
Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min. In Simple Western only 10-15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:50 The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Caspase-1

Caspase-1, also known as interleukin-1-beta (IL-1beta) -converting enzyme (ICE), is a member of the cysteine-aspartic protease (caspase) family (1). The caspase family are aspartic proteolytic enzymes that play a key role in apoptotic cell death and inflammation (1). Caspase-1 is synthesized as an inactive proenzyme with a theoretical molecular weight of approximately 45 kDa and is 404 amino acids (aa) in length. The caspase-1 protein structure contains an N-terminal CARD (caspase recruitment domain), a large 20kDa P20 subunit (aa 120-297), and a small 10kDa P10 subunit (aa 317-404) (1, 2). The proenzyme undergoes cleavage at aspartic acid (Asp) residues to produce the active caspase-1 enzyme which exists as a heterotetramer (homodimer of the P20 and P10 heterodimers) (2). Inflammasome complexes sense various microbial and endogenous signals, including ligand-receptor interaction and growth factor deprivation, which results in the processing of pro-caspase to active caspase-1 (3, 4).The active caspase-1 is responsible for the cleavage and maturation of pro-inflammatory cytokines IL-1beta and IL-18, and promotes an inflammatory form of cell death induced by bacterial pathogens called pyroptosis (3, 4). Mature IL-1beta has a role in the immune reaction and recruiting inflammatory cells to the site of infection while IL-18 plays a role in interferon-gamma production and promotes cytolytic activity in natural killer cells (4).

Given the role of IL-1beta in inflammation, it makes sense that many diseases and pathologies have been associated with dysregulation of caspase-1 activation and the inflammasome (3, 4). The inflammasome is a multiprotein complex comprised of Nod-like receptor (NLR) family members and the adapter ASC (apoptosis-associated speck-like protein containing a CARD) which are crucial for capase-1 activation (3-5). For instance, the neuronal apoptosis inhibitory protein (NAIP)/NLRC4 inflammasome has been associated with colorectal cancer, breast cancer, and glioma pathogenesis (5). Caspase-1 activation and mutations in the inflammasome have also been linked to Chron's disease and Alzheimer's disease (4). In addition to immune and inflammatory related disorder, the inflammasome has been linked to metabolic and obesity related disorders including diabetes and cardiovascular disease (6). Finally, caspase-1 deficient mice exhibit enhanced epithelial cell proliferation in the colon, increased tumor formation, and reduced apoptosis (1). A more thorough understanding of the inflammasome-caspase-1 signaling pathway will be important for understanding disease pathology and potential therapeutic development.

Alternative names for caspase-1 includes CASP1, CASP1 nirs variant 1, EC 3.4.22.36, ICE, IL-1 beta-converting enzyme, IL1BC, IL1BCE, IL1B-converstase, interleukin-1 beta convertase, and p45.

References

1. Shalini, S., Dorstyn, L., Dawar, S., & Kumar, S. (2015). Old, new and emerging functions of caspases. Cell death and differentiation. https://doi.org/10.1038/cdd.2014.216

2. Chang, H. Y., & Yang, X. (2000). Proteases for cell suicide: functions and regulation of caspases. Microbiology and molecular biology reviews: MMBR. https://doi.org/10.1128/mmbr.64.4.821-846.2000

3. Vanaja, S. K., Rathinam, V. A., & Fitzgerald, K. A. (2015). Mechanisms of inflammasome activation: recent advances and novel insights. Trends in cell biology. https://doi.org/10.1016/j.tcb.2014.12.009

4. Franchi, L., Eigenbrod, T., Munoz-Planillo, R., & Nunez, G. (2009). The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis. Nature immunology. https://doi.org/10.1038/ni.1703

5. Kay, C., Wang, R., Kirkby, M., & Man, S. M. (2020). Molecular mechanisms activating the NAIP-NLRC4 inflammasome: Implications in infectious disease, autoinflammation, and cancer. Immunological reviews. https://doi.org/10.1111/imr.12906

6. Pham, D., Park, P. (2020). Recent insights on modulation of inflammasomes by adipokines: a critical event for the pathogenesis of obesity and metabolism-associated diseases. Archives of Pharmacal Research. https://doi.org/10.1007/s12272-020-01274-7

Alternate Names

CASP1, Caspase1, ICE

Entrez Gene IDs

834 (Human); 12362 (Mouse); 25166 (Rat)

Gene Symbol

CASP1

UniProt

P29466 (Human)

Additional Caspase-1 Products

Product Documents for Caspase-1 Antibody (14F468) - Azide and BSA Free

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Product Specific Notices for Caspase-1 Antibody (14F468) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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