Anti-Influenza A Virus PA protein antibody [EPR28250-39]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells (human embryonic kidney epithelial cell) transfected with a Influenza A Virus PA protein expression vector containing a Myc-His tag cells labelling Influenza A Virus PA protein with ab313745 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Influenza A Virus PA protein with ab313745 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human kidney. The section was incubated with ab313745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Immunohistochemical analysis of paraffin-embedded Panel A HEK-293T tissue labeling Influenza A Virus PA protein with ab313745 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) HEK-293T transfected with a Influenza A Virus PA protein expression vector and no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab313745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Influenza A Virus PA protein with ab313745 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining in 293T cells transfected with a Influenza A Virus PA protein expression vector containing a myc tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 5ug/ml dilution (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 2 ug/mL dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Influenza A Virus PA protein with ab313745 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on mouse kidney. The section was incubated with ab313745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Influenza A Virus PA protein with ab313745 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on rat kidney. The section was incubated with ab313745 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• I-ELISA

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Indirect ELISA - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Indirect ELISA analysis of ab313745 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.
Antigen : Influenza A Virus PA protein.
Antigen concentration : 1000 ng/ml

• IP

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Immunoprecipitation - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Influenza A Virus PA protein was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with a Influenza A Virus PA protein expression vector containing a His-tag whole cell lysate with ab313745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313745 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : 293T (human embryonic kidney epithelial cell) cells transfected with a Influenza A Virus PA protein expression vector containing a His-tag whole cell lysate Lane 2 : 293T (human embryonic kidney epithelial cell) cells transfected with a Influenza A Virus PA protein expression vector containing a His-tag whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313745 in 293T whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 5 seconds

All lanes:

Immunoprecipitation - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (ab313745) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) cells transfected with a Influenza A Virus PA protein expression vector containing a His-tag whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 5s

• WB

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Western blot - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (AB313745)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The lower MW band at approximately 65kDa is expected to be a degradation product. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-Influenza A Virus PA protein antibody [EPR28250-39] (ab313745) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Influenza A Virus PA protein 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: 83 kDa

false

Exposure time: 5.5s