Anti-Caspase-1 antibody [EPR19672] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-Caspase-1 antibody [EPR19672] - BSA and Azide free (AB238979)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207802).

All lanes:

Western blot - Anti-Caspase-1 antibody [EPR19672] (<a href='/en-us/products/primary-antibodies/caspase-1-antibody-epr19672-ab207802'>ab207802</a>) 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

• IP

Lab

Immunoprecipitation - Anti-Caspase-1 antibody [EPR19672] - BSA and Azide free (AB238979)

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 1/1000 dilution .

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207802).

All lanes:

Immunoprecipitation - Anti-Caspase-1 antibody [EPR19672] (<a href='/en-us/products/primary-antibodies/caspase-1-antibody-epr19672-ab207802'>ab207802</a>)

Predicted band size: 45 kDa

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

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-Caspase-1 antibody [EPR19672] - BSA and Azide free (AB238979)

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] (<a href='/en-us/products/primary-antibodies/caspase-1-antibody-epr19672-ab207802'>ab207802</a>) 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

• WB

CiteAb

Western blot - Anti-Caspase-1 antibody [EPR19672] - BSA and Azide free (AB238979)

Caspase-1 western blot using anti-Caspase-1 antibody [EPR19672] - BSA and Azide free ab238979. Publication image and figure legend from Liu, J., Yao, L., et al., 2019, Aging (Albany NY), PubMed 31553952.

ab238979 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 ab238979 please see the product overview.

The influences of LncRNA-XIST on oxidative stress and pyroptosis of NSCLC cell lines. MDA levels was detected in (A) A549 cells and (C) H1299 cells. Extracellular NADPH oxidase-derived superoxide was detected in (B) A549 cells and (D) H1299 cells. DHE staining was used to detect ROS levels in (E) A549 cells, (F) H1299 cells and (G) HBE cells. (H, K) Western Blot was used to detect pyroptosis associated proteins (NLRP3, Cleaved Caspase-1 and IL-1β), which was normalized to β-actin and (I, L) quantified by Image J software. (J, M) ELSIA kit was used to detect IL-18 levels in A549 cell supernatants. (N, O) Colony formation assay was performed to detect cell proliferative abilities. (P) CCK-8 assay was used to detect A549 cell proliferation. "NAC" means N-acetyl cysteine treatment. "KD" means knock-down of LncRNA-XIST. "N+K" means NAC treatment plus knocking down LncRNA-XIST. ("NS" represented no statistical significance, "*" represented p < 0.05, "**" represented p < 0.01).

false