Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded 293T (human tissue labeling Influenza B virus Nucleoprotein with ab313885 at 1/2000 (0.254 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on (A) 293T transfected with an Influenza B virus NP expression vector and no staining on (B) 293T transfected with empty vector. The section was incubated with ab313885 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 B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Influenza B virus Nucleoprotein with ab313885 at 1/2000 (0.254 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human kidney. The section was incubated with ab313885 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 B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with an Influenza B virus NP gene expression vector containing a myc tag cells labelling Influenza B virus Nucleoprotein with ab313885 at 1/50 (10.16 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing nuclear staining in 293T cells transfected with an Influenza B virus NP gene expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain at 1/100 (0.38 µg/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 µg/ml) dilution.

• IP

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Immunoprecipitation - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation.

Influenza B virus Nucleoprotein was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with an Influenza B virus NP gene expression vector containing a myc-His-tag® whole cell lysate with ab313885 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313885 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 an Influenza B virus NP gene expression vector containing a myc-His-tag® whole cell lysate
Lane 2 : ab313885 IP in 293T (human embryonic kidney epithelial cell) cells transfected with an Influenza B virus NP gene expression vector containing a myc-His-tag® whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313885 in 293T cells transfected with an Influenza B virus NP gene expression vector containing a myc-His-tag® whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] (<a href='/en-us/products/primary-antibodies/influenza-b-virus-nucleoprotein-antibody-epr28205-27-ab313885'>ab313885</a>) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) cells transfected with an Influenza B virus NP gene expression vector containing a myc-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: 180s

• I-ELISA

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Indirect ELISA - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab313885 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 B virus Nucleoprotein. Antigen concentration : 1000 ng/ml.

• WB

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Western blot - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] - BSA and Azide free (AB313886)

This data was developed using ab313885, the same antibody clone in a different buffer formulation. 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 Exposure time : 26 seconds

All lanes:

Western blot - Anti-Influenza B virus Nucleoprotein antibody [EPR28205-27] (<a href='/en-us/products/primary-antibodies/influenza-b-virus-nucleoprotein-antibody-epr28205-27-ab313885'>ab313885</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 2:

293T cells transfected with an Influenza B virus NP gene expression vector containing a myc-His-tag® whole cell lysate at 5 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 1/20000 dilution

Observed band size: 62 kDa,65 kDa

false

Exposure time: 26s