Caspase-8 Antibody (90A992) - BSA Free

Novus Biologicals | Catalog # NB100-56527

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Primate

Cited:

Human, Insect - Drosophila melanogaster, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Simple Western, CyTOF-ready

Cited:

Western Blot, Simple Western, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 90A992

Format

BSA Free
View all Caspase-8 Primary Antibodies »

Product Specifications

Immunogen

The immunogen for this product falls between amino acids 360-385 of the human Caspase 8 protein.

Reactivity Notes

Chimpanzee and Rhesus Monkey reactivity noted.

Specificity

This can be used for detection of the pro-form of Caspase-8.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Caspase-8 Antibody (90A992) - BSA Free

Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Western blot analysis of Caspase-8 in Jurkat cells using Caspase-8 antibody at 1 ug/ml. Cells were treated with 2 uM staurosporine for different time periods. Caspase-8 activation is detected in western blots by the presence of Caspase-8 cleavage fragments. The antibody detected both pro (full length) and active (cleaved) Caspase-8, depending on the treatment time points. A basal level of endogenously cleaved Caspase-8 can be see in untreated Jurkat cells. Goat anti-mouse lg HRP secondary antibody and PicoTect ECL substrate solution were used for this test.
Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Caspase-8 expression in 786-O and A-498 cellsClose
Immunohistochemistry-Paraffin: Caspase-8 Antibody (90A992) - BSA Free [NB100-56527]

Immunohistochemistry-Paraffin: Caspase-8 Antibody (90A992) - BSA Free [NB100-56527]

Immunohistochemistry-Paraffin: Caspase-8 Antibody (90A992) [NB100-56527] - Formalin-fixed, paraffin-embedded human spleen (top) and breast (bottom) stained with Caspase-8 antibody at 4 ug/ml. Localization can be cytoplasmic and nuclear. Cancer/normal adjacent tissue array was used for this test. 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.
Flow Cytometry: Caspase-8 Antibody (90A992) - BSA Free [NB100-56527]

Flow Cytometry: Caspase-8 Antibody (90A992) - BSA Free [NB100-56527]

Flow Cytometry: Caspase-8 Antibody (90A992) [NB100-56527] - Flow cytometric analysis of Caspase-8 in HeLa cells using 0.1 ug of Caspase-8 antibody. Shaded histogram represents cells without antibody; green represents isotype control; red represents Caspase-8 antibody. Goat anti-mouse IgG-FITC secondary antibody was used for this test. IC-Flow (Intracellular Staining Flow Cytometry Kit) was used to fix and prepare the cells for staining.
Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Analysis using the Biotin conjugate of NB100-56527. Detection of human Caspase-8 using Jurkat lysates with NB100-55786 at 2 ug/ml (lane 1) and 0.5 ug/ml (lane 2) dilution. NB100-55786 only detects 55 kDa Caspase-8 in Jurkat cells.
Simple Western: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Simple Western: Caspase-8 Antibody (90A992)BSA Free [NB100-56527]

Simple Western: Caspase-8 Antibody (90A992) [NB100-56527] - Simple Western lane view shows a specific band for Caspase 8 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Caspase-8 Antibody (90A992) - BSA Free

Simple Western: Mouse Monoclonal Caspase-8 Antibody (90A992) [NB100-56527] -

Simple Western: Mouse Monoclonal Caspase-8 Antibody (90A992) [NB100-56527] - S2/IAPinh induces activation of the extrinsic apoptosis pathway via degradation of cIAP-1. (A) Schematic diagram of the intrinsic and extrinsic apoptosis pathways: arrows represent stimulation. TNFR, tumor necrosis factor receptor; DR4-5, death receptor 4–5; cIAP-1/2, cellular inhibitor of apoptosis protein-1/2; RIPK1, Receptor-interacting serine/threonine-protein kinase 1. (B) Protein expression of cIAP-1, cIAP-2, and XIAP in HPAC cells treated with vehicle, SW43 (10 µM), IAPinh (10 µM), or S2/IAPinh (10 µM) for 6 h. The precursor and cleaved forms of caspases 3, 8, and 9 were also analyzed for these cells using Wes automated capillary blotting system (Protein Simple). (C) Quantification of protein expression. Relative densitometry of each band normalized to the total protein. Data shown as means ± SEM. **P < 0.01, **** P < 0.0001. (D) Ratio of Caspase 3/7 counts to NucRed counts in HPAC cells treated with vehicle, SW43 (10 µM), IAPinh (10 µM), combination of SW43 (10 µM) and IAPinh (10 µM), or S2/IAPinh (10 µM) measured by the IncuCyte system (Sartorius). Bar graph shows the ratio of Caspase 3/7 at 48 h for each treatment. Data shown as means ± SEM. ****P < 0.001. (E) Activity of cell death in AsPC-1 cells was measured using YOYO-1 iodide on the IncuCyte (Sartorius). Representative images of AsPC-1 cells treated with or without Z-VAD-FMK and S2/IAPinh (10 uM) at baseline and 36 h after treatment. Scale bars are equal to 20 µm. (F) The AUC of lethal fraction at 36 h. Data shown as means ± SEM. ****P < 0.0001. Image collected and cropped by CiteAb under a CC-BY license from the following publication: The novel drug candidate S2/IAPinh improves survival in models of pancreatic and ovarian cancer. Sci Rep (2024). Not internally tested by Novus Biologicals.
Caspase-8 Antibody (90A992) - BSA Free

Simple Western: Mouse Monoclonal Caspase-8 Antibody (90A992) [NB100-56527] -

Simple Western: Mouse Monoclonal Caspase-8 Antibody (90A992) [NB100-56527] - S2/IAPinh induces tumor cell death via activation of the extrinsic apoptosis pathway in vivo. KP2 tumor samples collected from animals at 48 h after the treatment with either vehicle, SW43, IAPinh, or S2/IAPinh, were used for analyses. (A) Representative images of TUNEL labeled apoptosis cells in KP2 tumor samples of each group. Nuclei were stained in blue with Hoechst, TUNEL positive cells are in red. Scale bars are equal to 20 µm. (B) Quantification of TUNEL positive cells per area in each group. Data are shown as means ± SEM; ****P < 0.0001. (C) Protein expression of cIAP-1, cIAP-2, and XIAP in KP2 tumor samples of each group. The precursor and cleaved forms of caspases 3, 8, and 9 were also analyzed for these cells using Wes automated capillary blotting system (Protein Simple). (D) Quantification of protein expression. Relative densitometry of each band normalized to the total protein. Data shown as means ± SEM. *P < 0.05, **P < 0.01, **** P < 0.0001. Image collected and cropped by CiteAb under a CC-BY license from the following publication: The novel drug candidate S2/IAPinh improves survival in models of pancreatic and ovarian cancer. Sci Rep (2024). Not internally tested by Novus Biologicals.

Applications for Caspase-8 Antibody (90A992) - BSA Free

Application
Recommended Usage

Flow Cytometry

0.1-0.5 ug/ml

Immunohistochemistry

reported in scientific literature (PMID 31034097)

Immunohistochemistry-Paraffin

4 ug/ml

Simple Western

1:100

Western Blot

0.5-2 ug/ml
Application Notes
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: Tested in Hek293 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:100, apparent MW was 16 kDa.This antibody is CyTOF ready.

Reviewed Applications

Read 1 review rated 4 using NB100-56527 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

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-8

Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Caspase-8 (FLICE) forms a direct link between the activation of CD95 and the caspase pathway. Overexpression of Caspase-8 induces apoptosis, which can be blocked by inhibitors specific for the ICE family.

Alternate Names

CASP8, Caspase8, Mch5

Gene Symbol

CASP8

Additional Caspase-8 Products

Product Documents for Caspase-8 Antibody (90A992) - BSA Free

Certificate of Analysis

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Product Specific Notices for Caspase-8 Antibody (90A992) - 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.

Citations for Caspase-8 Antibody (90A992) - BSA Free

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Customer Images

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: 786-O and A-498 whole cell lysates
    Species: Human
    Verified Customer | Posted 04/29/2015
    Caspase-8 expression in 786-O and A-498 cells
    Caspase-8 Antibody (90A992) - BSA Free NB100-56527

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Protocols

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FAQs for Caspase-8 Antibody (90A992) - BSA Free

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  • Q: May we ask if NB600-1343, NB100-56527 and NB500-208 can recognize both active form and profor

    A: Product NB100-56527 recognises both pro and active forms of Caspase 8. NB500-208 recognises only the active form, and NB600-1343 has not been tested for specificity at this time.

  • Q: May we ask the MW difference between the active form and the proform of Caspase 8 while performing WB?

    A: In treated cells induced to undergo apoptosis, caspase-8 migrates as a 55/53 kDa (pro-form), 41/42 kDa (a cleaved/active or intermediate form), and 18 kDa (active form). The proform (55/53 kDa) is still seen in treated cells because not all cells undergo apoptosis at once.

  • Q: May we ask if NB600-1343, NB100-56527 and NB500-208 can recognize both active form and profor

    A: Product NB100-56527 recognises both pro and active forms of Caspase 8. NB500-208 recognises only the active form, and NB600-1343 has not been tested for specificity at this time.

  • Q: May we ask the MW difference between the active form and the proform of Caspase 8 while performing WB?

    A: In treated cells induced to undergo apoptosis, caspase-8 migrates as a 55/53 kDa (pro-form), 41/42 kDa (a cleaved/active or intermediate form), and 18 kDa (active form). The proform (55/53 kDa) is still seen in treated cells because not all cells undergo apoptosis at once.

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Associated Pathways

Apoptosis Signaling Pathway Apoptosis Signaling Pathway Thumbnail