Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human |
Applications: | WB, IF-Cell, IHC-P, FC |
Clonality: | Monoclonal |
Clone number: | PSH04-77 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Concentration: | 1ug/ul |
Purification: | Protein A affinity purified. |
Molecular weight: | Predicted band size: 55 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide within human Caspase-8 aa 325-374 / 479. |
Positive control: | Jurkat cell lysate, Jurkat treated with 25μM Etoposide for 5 hours cell lysate, Jurkat treated with 25μM Etoposide for 16 hours cell lysate, Jurkat treated with 1μM staurosporine for 3 hours cell lysate, HeLa cell lysate, HeLa treated with 1μM staurosporine for 3 hours cell lysate, HeLa treated with 100μM Etoposide for 4 hours cell lysate, human tonsil tissue, human liver tissue, HeLa cells treated with 1μM staurosporine for 3 hours. |
Subcellular location: | Cytoplasm, Nucleus. |
Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:2,000 1:100 1:200-1:1,000 1:1,000 |
Uniprot #: | SwissProt: Q14790 Human |
Alternative names: | ALPS2B Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein Apoptotic cysteine protease Apoptotic protease Mch-5 Apoptotic protease Mch5 CAP4 CASP-8 CASP8 CASP8_HUMAN Caspase8 Caspase 8 Caspase 8 apoptosis related cysteine peptidase Caspase-8 subunit p10 CED 3 FADD Like ICE FADD-homologous ICE/CED-3-like protease FADD-like ICE FLICE FLJ17672 ICE-like apoptotic protease 5 MACH alpha 1/2/3 protein MACH MACH beta 1/2/3/4 protein MCH5 MGC78473 MORT1 associated ced 3 homolog MORT1-associated CED-3 homolog OTTHUMP00000163717 OTTHUMP00000163720 OTTHUMP00000163724 OTTHUMP00000163725 OTTHUMP00000165062 OTTHUMP00000165063 OTTHUMP00000165064 OTTHUMP00000206552 OTTHUMP00000206582 |
Fig1:
Western blot analysis of Caspase-8 on different lysates with Rabbit anti-Caspase-8 antibody (HA722181) at 1/2,000 dilution. Lane 1: Jurkat cell lysate Lane 2: Jurkat treated with 25μM Etoposide for 5 hours cell lysate Lane 3: Jurkat treated with 25μM Etoposide for 16 hours cell lysate Lane 4: Jurkat treated with 1μM staurosporine for 3 hours cell lysate Lane 5: HeLa cell lysate Lane 6: HeLa treated with 1μM staurosporine for 3 hours cell lysate Lane 7: HeLa treated with 100μM Etoposide for 4 hours cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 55 kDa Observed band size: 18-55 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722181) at 1/2,000 dilution was used in antibody diluent at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HeLa cells treated with or without 1μM staurosporine for 3 hours labeling Caspase-8 with Rabbit anti-Caspase-8 antibody (HA722181) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Caspase-8 antibody (HA722181) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
Fig3:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Caspase-8 antibody (HA722181) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722181) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Caspase-8 antibody (HA722181) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722181) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig5:
Flow cytometric analysis of HeLa cells treated with or without 1μM staurosporine for 3 hours cells labeling Caspase-8. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722181, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |