Anti-IL-26 antibody [EPR22268-141]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-IL-26 antibody [EPR22268-141] (AB224198)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia cell line) cells labeling IL-26 with ab224198 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in THP-1 cell line treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 16h, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red). Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Cells were either untreated, or treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 16h, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-26 antibody [EPR22268-141] (AB224198)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling IL-26 with ab224198 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of human colon (PMID : 18483078, 28852311) is observed. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. The section was incubated with ab224198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-26 antibody [EPR22268-141] (AB224198)

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue labeling IL-26 with ab224198 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of human cervical carcinoma (PMID : 18483078, 28852311) is observed. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. The section was incubated with ab224198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IP

Unknown

Immunoprecipitation - Anti-IL-26 antibody [EPR22268-141] (AB224198)

IL-26 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) (treated with 80nM PMA (Phorbol-12-myristate-13-acetate) overnight, then 100ng/ml LPS (lipopolysaccharide) for 3h and add 300ng/ml BFA (Brefeldin A) for 3h) whole cell lysate with ab224198 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224198 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

Lane 1 : THP-1 (treated as above) whole cell lysate 10 μg (Input).
Lane 2 : ab224198 IP in THP-1 (treated as above) whole cell lysate.
Lane 3 : : Rabbit monoclonal IgG (ab172730) instead of ab224198 in THP-1 (treated as above) whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 3 mins.

All lanes:

Immunoprecipitation - Anti-IL-26 antibody [EPR22268-141] (ab224198)

Predicted band size: 20 kDa

Observed band size: 27 kDa

false

• WB

Unknown

Western blot - Anti-IL-26 antibody [EPR22268-141] (AB224198)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 14764663).

This antibody reacts with an unidentifiable protein around 75 kDa.

All lanes:

Western blot - Anti-IL-26 antibody [EPR22268-141] (ab224198) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate at 20 µg

Lane 2:

THP-1 treated with 80nM PMA (Phorbol-12-myristate-13-acetate) for 24 hours, then 1μg/ml LPS (lipopolysaccharides) for 24 hours, 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 100000 mg/mL

Predicted band size: 20 kDa

Observed band size: 27 kDa

false

Exposure time: 92s

• WB

Unknown

Western blot - Anti-IL-26 antibody [EPR22268-141] (AB224198)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IL-26 antibody [EPR22268-141] (ab224198) at 1/1000 dilution

All lanes:

His-tagged human IL-26 recombinant protein (aa 22-171), 20 ng

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: 20 kDa

false

Exposure time: 26s