Anti-Influenza A virus PB1 protein antibody [F5-19]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Influenza A virus PB1 protein antibody [F5-19] (AB324448)

Confocal image showing positive staining in 293T cells transfected with an influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
A : 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-his-tag
B : 293T cells transfected with an influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag
C : 293T cells transfected with an influenza B virus PB1 expression vector containing a myc-his-tag
D : 293T cells transfected with an influenza C virus PB1 expression vector containing a myc-his-tag

Primary antibody ab324448 used at a 1/100 (9.46 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green).

ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 (2 ug/ml) dilution (Magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100.

• IP

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Immunoprecipitation - Anti-Influenza A virus PB1 protein antibody [F5-19] (AB324448)

Influenza A virus PB1 protein was immunoprecipitated from 0.35 mg 293T cells transfected with a influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag whole cell lysate with ab324448 at 1/50 dilution (2 ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324448 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Influenza A virus PB1 protein antibody [F5-19] (ab324448) at 1/1000 dilution

Lane 1:

(Input) 293T cells transfected with a influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag whole cell lysate at 10 µg

Lane 2:

ab324448 at 1/50 IP in 293T cells transfected with a influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag whole cell lysate at 10 µg

Lane 3:

Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control (<a href='/en-us/products/primary-antibodies/mouse-igg1-kappa-monoclonal-mopc-21-isotype-control-ab18443'>ab18443</a>) instead of ab324448 in 293T cells transfected with a influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - Anti-mouse IgG for IP (HRP) (<a href='/en-us/products/secondary-antibodies/mouse-igg-for-ip-hrp-ab131368'>ab131368</a>) at 1/5000 dilution

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Influenza A virus PB1 protein antibody [F5-19] (AB324448)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Influenza A virus PB1 protein antibody [F5-19] (ab324448) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with an influenza A virus (H3N2) PB1 expression vector containing a myc-his-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with an influenza A virus (H3N2) PB2 expression vector containing a myc-his-tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with an influenza B virus PB1 expression vector containing a myc-his-tag whole cell lysate at 20 µg

Lane 5:

293T cells transfected with an influenza C virus PB1 expression vector containing a myc-his-tag whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

false

Exposure time: 6s