Anti-Caspase-1 antibody [EPR19672]

• WB

Lab

Western blot - Anti-Caspase-1 antibody [EPR19672] (AB207802)

False colour image of Western blot : Anti-Caspase-1 antibody [EPR19672] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab207802 was shown to bind specifically to Caspase-1. A band was observed at 22, 35-40, 48 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP1 knockout cell line ab276116 (knockout cell lysate ab277988). To generate this image, wild-type and CASP1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Caspase-1 antibody [EPR19672] (ab207802) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 40 µg

Lane 2:

CASP1 knockout THP-1 cell lysate at 40 µg

Lane 2:

Western blot - Human CASP1 (Caspase-1) knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-casp1-caspase-1-knockout-thp-1-cell-line-ab276116'>ab276116</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 35-40 kDa,48 kDa

false

• IP

Lab

Immunoprecipitation - Anti-Caspase-1 antibody [EPR19672] (AB207802)

ab207802 Immunoprecipitating Caspase-1 in THP-1 (human acute monocytic leukemia) whole cell lysate. 2μg of capture antibody in 0.35mg lysate was used. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/1000 and VeriBlot for IP Detection Reagent (HRP) ab131366 was used for detection at a dilution of 1/1000.

Lane 1 : THP-1 (human acute monocytic leukemia ) whole cell lysate
Lane 2 : THP-1 (human acute monocytic leukemia ) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207802 in THP-1 (human acute monocytic leukemia ) whole cell lysate

Blocking buffer : 5% NFDM/TBST

Diluting buffer : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Caspase-1 antibody [EPR19672] (ab207802)

Predicted band size: 45 kDa

Observed band size: 20 kDa,22 kDa,33-35 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Caspase-1 antibody [EPR19672] (AB207802)

Blocking/Dilution buffer : 5% NFDM/TBST.

There are six pro-caspase 1 isoforms, generating products between 30-45kDa, Pro-Caspase 1 is cleaved into p20(p22) and p10(p12) subunits. PMID : 8662843. PMID : 7595215. The product recognizes p20(p22) cleavage fragment where the immunogen is located.

All lanes:

Western blot - Anti-Caspase-1 antibody [EPR19672] (ab207802) at 1/1000 dilution

Lane 1:

Untreated THP-1 (Human monocytic leukemia cell line) whole cell lysate at 10 µg

Lane 2:

THP-1 (Human monocytic leukemia cell line) treated with 80 nM TPA overnight, then treated with 1 μg/ml LPS for 8 hours whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 20 kDa,22 kDa,42 kDa,45 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Caspase-1 antibody [EPR19672] (AB207802)

Blocking/Dilution buffer : 5% NFDM/TBST.

Pro-Caspase 1 is cleaved into p20(p22) and p10(p12) subunits. PMID : 8662843. PMID : 7595215. The product recognizes p20(p22) cleavage fragment where the immunogen is located.

The 37kDa band may also include isoform 5, but we have no experimental data to verify this.

All lanes:

Western blot - Anti-Caspase-1 antibody [EPR19672] (ab207802) at 1/1000 dilution

Lane 1:

Human lymph node lysate at 10 µg

Lane 2:

Human thymus lysate at 10 µg

Lane 3:

Human spleen lysate at 10 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 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 22 kDa,35 kDa,45 kDa

false

Exposure time: 1min

• WB

CiteAb

Western blot - Anti-Caspase-1 antibody [EPR19672] (AB207802)

Caspase-1 western blot using anti-Caspase-1 antibody [EPR19672] ab207802. Publication image and figure legend from Yuan, R., He, J., et al., 2020, J Immunol Res, PubMed 33344657.

ab207802 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 ab207802 please see the product overview.

B4 suppressed NLRP3 inflammasome activation. (a, b) J774A.1 cells and THP-1 cells were pretreated with B4 for 4 h, then cocultured with LPS (1 μg/mL) for another 5 h, and stimulated with ATP (5 mM) for 30 min. The expression levels of NLRP3, caspase-1, and IL-1β were determined by Western blot. All the experiments were performed three times independently. **p < 0.01 and ***p < 0.001 versus LPS+ATP group.

false