Anti-IL-22 antibody [EPR20211-49] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IL-22 antibody [EPR20211-49] - BSA and Azide free (AB263954)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell) transfected with myc-tagged IL-22 expression vector cells labelling IL-22 with ab227033 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (left)and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227033).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IL-22 antibody [EPR20211-49] - BSA and Azide free (AB263954)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling IL-22 with ab227033 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and staining in 293T cells transfected with myc-tagged IL-22 expression vector. The vector also contains a PDGFR transmembrane domain. is observed. ab223894 Anti-Myc tag antibody [9E10] (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227033).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IL-22 antibody [EPR20211-49] - BSA and Azide free (AB263954)

Intracellular flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Th17 polarized human PBMC treated with 50ng/ml PMA (ab120297), 200ng/ml Ionomycin calcium (ab120116) and 300ng/ml BFA (ab193369) for 4h cells labelling IL-22 with ab227033 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) Left isotype control (left)and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227033).

• IP

Lab

Immunoprecipitation - Anti-IL-22 antibody [EPR20211-49] - BSA and Azide free (AB263954)

IL-22 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) transfected with myc-tagged IL-22 expression vector whole cell lysate with ab227033 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab227033 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with myc-tagged IL-22 expression vector whole cell lysate 10ug

Lane 2 : ab227033 IP in 293T transfected with myc-tagged IL-22 expression vector whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227033 in 293T transfected with myc-tagged IL-22 expression vector whole cell lysate

The vector containing a PDGFR transmembrane domain (~6 kDa). The molecular weight observed is consistent with what has been described in the literature (PMID : 26059044)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 20 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227033).

All lanes:

Immunoprecipitation - Anti-IL-22 antibody [EPR20211-49] (<a href='/en-us/products/primary-antibodies/il-22-antibody-epr20211-49-ab227033'>ab227033</a>)

Predicted band size: 20 kDa

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