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<strong> Figure 1 WB Validation in Human Tissues </strong><br>Loading: 15 μg of lysate Antibodies: Caspase-8, 3473, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 WB Validation in Mouse 3T3/NIH Cells</strong><br>Loading: 15 μg of lysate Antibodies: Caspase-8, 3473, 1 μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong> Figure 3 WB Validation in Rat Testis </strong><br>Loading: 15 μg of lysate Antibodies: Caspase-8, 3473, 2 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of Caspase-8 in Human Liver</strong><br> Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Caspase-8 antibody (3473) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunohistochemistry Validation of Caspase-8 in Mouse Liver</strong><br> Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Caspase-8 antibody (3473) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunohistochemistry Validation of Caspase-8 in Rat Liver</strong><br> Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-Caspase-8 antibody (3473) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunofluorescence Validation of Caspase-8 in Human Spleen</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human spleen tissue labeling Caspase-8 with 3473 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
<strong> Figure 8 WB Validation in Human K562 Cells </strong><br>Loading: 15 μg of lysate Antibodies: Caspase-8, 3473, 1 μ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
Figure 1 WB Validation in Human Tissues
Loading: 15 μg of lysate Antibodies: Caspase-8, 3473, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

Caspase-8 Antibody

CATALOG NUMBER: 3473

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB, IHC-P/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-Caspase-8 antibody (3473) was raised against a peptide corresponding to 16 amino acid synthetic peptide near the carboxyl terminus of human Caspase-8.

The immunogen is located within the last 50 amino acids of Caspase-8.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB, IHC-P/IF
Application Note:
WB: 1-2 μg/mL; IHC-P: 1-2 μg/mL; IHC-P/IF: 10-20 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Specificity:
Depending on cell lines or tissues used, either full-length or other cleavage products may be observed.
Positive Control 3:
Cat. No. 1466 - Rat Spleen Tissue Lysate
Predicted Molecular Weight:
Predicted: 55 kD

Observed: 55 kD
Purification:
Caspase-8 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
Caspase-8 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
Caspase-8 antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
CASP8
Additional Names:
Caspase-8 Antibody: CAP4, MACH, MCH5, FLICE, ALPS2B, Casp-8, Caspase-8, Apoptotic cysteine protease, CASP-8
Protein Accession Number:
NP_001219
Protein Gi Number:
15718704
Ncbi Gene Id Number:
841
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
Caspase-8 Antibody: Caspases are a family of cysteine proteases that can be divided into the apoptotic and inflammatory caspase subfamilies. Unlike the apoptotic caspases, members of the inflammatory subfamily are generally not involved in cell death but are associated with the immune response to microbial pathogens. The apoptotic subfamily can be further divided into initiator caspases, which are activated in response to death signals, and executioner caspases, which are activated by the initiator caspases and are responsible for cleavage of cellular substrates that ultimately lead to cell death. Caspase-8 is an initiator caspase that was identified as a member of the Fas/APO-1 death-inducing signaling complex. The adaptor molecule FADD couples procaspase-8 to the Fas receptor death domain; subsequent oligomerization promotes procaspase-8 autoactivation. FLIP, a catalytically inactive caspase-8-like molecule inhibits these interactions and thus can inhibit apoptosis.
Background Reference 1:
Martinon F and Tschopp J. Inflammatory caspases: linking an intracellular innate immune system to autoinflammatory diseases. Cell 2004; 117:561-74.
Background Reference 2:
Zhivotovsky B and Orrenius S. Caspase-2 function in response to DNA damage. Biochim. Biophys. Res. Comm. 2005; 331:859-67.
Background Reference 3:
Wolf BB and Green DR. Suicidal tendencies: apoptotic cell death by caspase family proteinases. J. Biol. Chem. 1999; 274:20049-52.
Background Reference 4:
Muzio M, Chinnaiyan AM, Kischkel FC, et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 1996; 85:817-27.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

3473

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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