Anti-IL-18 antibody [EPR19954-188]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on Kupffer cells in human liver (PMID : 19084941). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on subsets of macrophages in tonsil (PMID : 28842466). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and PC-3 (human prostate adenocarcinoma cell line) (right panel) cells labeling IL-18 with ab243091 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : Jurkat (PMID 15086390).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on distal convoluted tubules of human kidney (PMID : 17687255). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Lab

Western blot - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Lanes 1-4 : Merged signal (red and green). Green - ab243091 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.

ab243091 Anti-IL-18 antibody [EPR19954-188] was shown to specifically react with IL-18 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265274 (knockout cell lysate ab256952) was used. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. ab243091 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

IL18 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human IL18 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-il18-knockout-hela-cell-line-ab265274'>ab265274</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

• WB

Supplier Data

Western blot - Anti-IL-18 antibody [EPR19954-188] (AB243091)

Exposure time : Lanes 1 & 2 : 3 minutes. Lane 3 : 26 seconds.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 3:

HaCaT (Human keratinocyte cell line) whole cell lysate at 20 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: 22 kDa

Observed band size: 14 kDa,22 kDa

false