Anti-HPV18 E7 antibody [EPR26911-29]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype control / HEK-293T cells transfected with Myc-tagged HPV18 Protein E7 overexpression construct / HPV45 Protein E7 overexpression construct / HPV70 Protein E7 overexpression construct / HPV39 Protein E7 overexpression construct / HPV16 Protein E7 overexpression construct / HPV18 Protein E6 overexpression construct labelling HPV18 E7 with ab308296 at 1/500 dilution (0.1 ug).
A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling HPV18 E7 with ab308296 at 1/100 dilution (5.09 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic and nuclear staining in HeLa cell line. Negative control : U-2 OS (PMID : 22135254). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-2 OS (human bone osteosarcoma epithelial cell) (Left panel) compared to HeLa (human cervical adenocarcinoma epithelial cell) (Right panel) cells labelling HPV18 E7 with ab308296 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

HPV18 E7 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab308296 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308296 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa whole cell lysate 10 ug Lane 2 : ab308296 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308296 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-HPV18 E7 antibody [EPR26911-29] (ab308296) at 1/30 dilution

Lanes 1 - 2:

ab308296 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab308296 in HeLa whole cell lysate

Observed band size: 17 kDa

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• WB

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Western blot - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : U-2 OS (PMID : 22135254), human testis, human stomach. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200, 000 dilution. Exposure time : 48 seconds.

All lanes:

Western blot - Anti-HPV18 E7 antibody [EPR26911-29] (ab308296) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

U-2 OS whole cell lysate at 20 µg

Lane 3:

Human testis tissue lysate at 20 µg

Lane 4:

Human stomach tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 17 kDa

false

• WB

Supplier Data

Western blot - Anti-HPV18 E7 antibody [EPR26911-29] (AB308296)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Weak cross-reactivity was observed with HPV45 E7 protein. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 26 seconds.

All lanes:

Western blot - Anti-HPV18 E7 antibody [EPR26911-29] (ab308296) at 1/1000 dilution

Lane 1:

293T cells transfected with a HPV18 E7 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a HPV45 E7 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a HPV39 E7 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a HPV16 E7 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a HPV18 E6 expression vector containing a His-tag 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: 17 kDa

false