Anti-Vaccinia Virus antibody [EPR28236-80]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Vaccinia Virus with ab312854 at 1/50 (10.28 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 Virion membrane protein A21 expression vector containing a GFP-tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labelling Vaccinia Virus with ab312854 at 1/100 (5.14 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) HEK-293T transfected with a Virion membrane protein A21 expression vector containing a GFP-tag cell pellet; no staining on (B) HEK-293T transfected with an empty vector containing a GFP-tag cell pellet. The section was incubated with ab312854 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-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Vaccinia Virus with ab312854 at 1/100 (5.14 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 ab312854 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with a Virion membrane protein A21 expression vector containing a GFP-tag (middle) / HEK-293T cells transfected with a empty expression vector containing a GFP-tag (Right) cells labelling Vaccinia Virus with ab312854 at 1/500 dilution (0.1 ug)/ Middle and 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® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling Vaccinia Virus with ab312854 at 1/100 (5.14 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 ab312854 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-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling Vaccinia Virus with ab312854 at 1/100 (5.14 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 ab312854 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-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Indirect ELISA analysis of ab312854 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 : Virion membrane protein A21.

Antigen concentration : 1000 ng/ml

• IP

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Immunoprecipitation - Anti-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Vaccinia Virus was immunoprecipitated from 0.35 mg 293T transfected with a Virion membrane protein A21 expression vector containing a GFP tag whole cell lysate with ab312854 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab312854 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : 293T transfected with a Virion membrane protein A21 expression vector containing a GFP tag whole cell lysate Lane 2 : 293T transfected with a Virion membrane protein A21 expression vector containing a GFP tag whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab312854 in 293T transfected with a Virion membrane protein A21 vector containing a GFP tag whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 58 seconds

All lanes:

Immunoprecipitation - Anti-Vaccinia Virus antibody [EPR28236-80] (ab312854) at 1/30 dilution

All lanes:

293T transfected with a Virion membrane protein A21 expression vector containing a GFP tag whole cell lysate at 1 µg

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: 58s

• WB

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Western blot - Anti-Vaccinia Virus antibody [EPR28236-80] (AB312854)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-GFP tag antibody (ab32146) staining at 1/1000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-Vaccinia Virus antibody [EPR28236-80] (ab312854) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a GFP tag whole cell lysate at 20 µg

Lane 2:

293T transfected with a Virion membrane protein A21 expression vector containi a GFP tag whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Human kidney tissue lysate at 20 µg

Lane 6:

Mouse kidney tissue lysate at 20 µg

Lane 7:

Rat kidney tissue 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: 46 kDa

false

Exposure time: 10s