Anti-IL-4I1/LAO antibody [EPR22070]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue stained for IL-4I1/LAO with ab222102 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Granular cytoplasmic staining in macrophages of human tonsil (PMID : 17356132; PMID : 19436310) is observed. Counterstained 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 was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilzed HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling IL-4I1/LAO (white) with ab222102 at 1/50 dilution, followed by ab150079 AlexaFluor^®647 Goat anti-Rabbit secondary at 1/1000 dilution. Confocal image showing cytoplasmic staining in HEK-293T cells transfected with GFP-tagged IL-4I1/LAO expression vector. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150079 AlexaFluor^®647 Goat anti-Rabbit secondary at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanolpermeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag labeling IL-4I1/LAO with ab222102at 1/500 dilution (Right) compared to Rabbit monoclonal IgG (ab172730) (Left) isotype control. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/2000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Immunohistochemical analysis of paraffin-embedded human difuse large B-cell lymphoma tissue stained for IL-4I1/LAO with ab222102 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Granular cytoplasmic staining in human diffuse large B-cell lymphoma (PMID : 19436310) is observed. Counterstained 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 was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Immunohistochemical analysis of paraffin-embedded human Hodgkin's lymphoma tissue stained for IL-4I1/LAO with ab222102 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Granular cytoplasmic staining in human Hodgkin's lymphoma (PMID : 19436310) is observed. Counterstained 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 was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

IL-4I1/LAO was immunoprecipitated from 0.35mg HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag whole cell lysate with ab222102 at 1/30 dilution. Western blot was perfromed from the immunoprecipitate using ab222102 at 1/1000 dilution, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : HEK-293T transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag whole cell lysate 10μg (Input).

Lane 2 : ab222102 IP in HEK-293 transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag whole cell lysate (+).

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab222102 in HEK-293T transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag whole cell lysate (-).

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 seconds.

All lanes:

Immunoprecipitation - Anti-IL-4I1/LAO antibody [EPR22070] (ab222102)

Predicted band size: 63 kDa

Observed band size: 100 kDa

false

• WB

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Western blot - Anti-IL-4I1/LAO antibody [EPR22070] (AB222102)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IL-4I1/LAO antibody [EPR22070] (ab222102) at 1/1000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (vector control), containing a myc-GFP-tag, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with IL-4I1/LAO expression vector containing a myc-GFP-tag, 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: 63 kDa

Observed band size: 100 kDa

false

Exposure time: 6s