Anti-E. coli antibody [EPR28234-47]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-E. coli antibody [EPR28234-47] (AB326069)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) co-cultured with EGFP expressing E. coli BL21(DE3) bacteria for 1.5 hours (Lower left and right) Untreated RAW 264.7 (Upper left and right) cells labelling E. coli with ab326069 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells with GFP-tag®

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-E. coli antibody [EPR28234-47] (AB326069)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) RAW 264.7 co-cultured with EGFP expressing E. coli BL21(DE3) bacteria for 1.5 hours cells labelling E. coli with ab326069 at 1/500 (1.01 µg/ml) dilution , followed by ab150083 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083) antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in RAW 264.7 co-cultured with EGFP expressing E. coli BL21(DE3) bacteria for 1.5 hours (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

was used to counterstain tubulin at 1/ dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150083 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083) at 1/1000 dilution.

• I-ELISA

Lab

Indirect ELISA - Anti-E. coli antibody [EPR28234-47] (AB326069)

Indirect ELISA analysis of ab326069 at 1000-0 ng/ml. The Secondary antibody used was Peroxidase AffiniPure Goat Anti-Rabbit IgG, Fc fragment specific at 1 : 20000 dilution dilution.

Antigen : E. coli.

Antigen concentration : 10^9 cell/ml

• WB

Lab

Western blot - Anti-E. coli antibody [EPR28234-47] (AB326069)

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

All lanes:

Western blot - Anti-E. coli antibody [EPR28234-47] (ab326069) at 1/1000 dilution

All lanes:

E. coli BL21(DE3) 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

Observed band size: 15-37 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-E. coli antibody [EPR28234-47] (AB326069)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-E. coli antibody [EPR28234-47] (ab326069) at 1/1000 dilution

Lane 1:

Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

RAW264.7 co‑cultured with E. coli (TOP10) bacteria for 1.5 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 1/100000 dilution

Observed band size: 15-37 kDa,36 kDa

false

Exposure time: 26s