Anti-Caspase-5 antibody [EP876Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-5 antibody [EP876Y] (AB40887)

Stainign of Caspase 5 on paraffin-embedded human breast carcinoma sections using ab40557 at a 1/50 dilution. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-Caspase-5 antibody [EP876Y] (AB40887)

Purified ab40887 at 1/50 dilution (2μg) immunoprecipitating Caspase-5 in HeLa treated with heat shock (42°C 1h) then recovery for 6h whole cell lysate.
Blocking and diluting Buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Caspase-5 antibody [EP876Y] (ab40887)

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with heat shock (42°C 1h) then recovery for 6h whole cell lysate (Input) at 10 µg

Lane 2:

ab40887 + HeLa treated with heat shock (42°C, 1h) then recovery for 6h whole cell lysate

Lane 3:

<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> instead of ab40887 in HeLa treated with heat shock (42&deg;C 1h) then recovery for 6h whole cell lysate (-)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 50 kDa

Observed band size: 47 kDa

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• WB

Unknown

Western blot - Anti-Caspase-5 antibody [EP876Y] (AB40887)

The cell lysates were prepared in 1%SDS Hot lysis method.

All lanes:

Western blot - Anti-Caspase-5 antibody [EP876Y] (ab40887) at 1/10000 dilution

All lanes:

HeLa cell lysate at 10 µg

Predicted band size: 50 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Anti-Caspase-5 antibody [EP876Y] (AB40887)

Blocking/Diluting Buffer and concentration :
5% NFDM/TBST

All lanes:

Western blot - Anti-Caspase-5 antibody [EP876Y] (ab40887) at 1/1000 dilution

All lanes:

HT-1080 (Human fibrosarcoma epithelial cell) prepared in 1%SDS Hot lysis method whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 47 kDa

false

• WB

CiteAb

Western blot - Anti-Caspase-5 antibody [EP876Y] (AB40887)

Caspase-5 western blot using anti-Caspase-5 antibody [EP876Y] ab40887. Publication image and figure legend from Zhang, Z., Shao, X., et al., 2018, Cell Death Dis, PubMed 30250284.

ab40887 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab40887 please see the product overview.

Iohexol induces TEC pyroptosis via activation of inflammatory caspases in mice and humans.a ELISA detection of mature IL-1β in human TEC culture supernatants 72 h after TECs treated with iohexol in a dose-dependent manner. b Immunoblot analysis of the inflammatory caspase-11 in mouse TECs and the inflammatory caspases 4/5 in human TECs with extracellular iohexol (20 mg/ml) or isosmotic mannitol for 72 h. c Quantification of the inflammatory caspases expression shows significant upregulation of caspase-11 and caspases 4/5 in mouse and human TECs, respectively. Statistics obtained from ANOVA. d mRNA expression of inflammatory caspases was determined by real-time PCR in human TECs after a 72 h period of iohexol (20 mg/ml) or isosmotic mannitol incubation. e Release of LDH showed that iohexol (20 mg/ml) or isosmotic mannitol incubation for 16 h led to marked cell lysis. Cell lysis was blocked by the pan-caspase inhibitor Z-VAD-FMK. f Incubation with iohexol (20 mg/ml) or isosmotic mannitol for 72 h increased mature IL-1β secretion, measured by ELISA in cell culture supernatants, whereas the response was abrogated by the pan-caspase inhibitor Z-VAD-FMK. Experiments were performed in triplicate. Ctrl, control. **p < 0.01, ***p < 0.001 vs. control or as indicated. Data are means ± SEM. Statistics obtained from ANOVA

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