Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] - BSA and Azide free (AB284054)

Intracellular Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a SARS-CoV-2 orf8 (Left) expression vector containing a myc tag or SARS-CoV2 orf3d (Right) expression vector containing GFP tag labeling SARS-CoV-2 orf3d with ab284040 at 1/500 dilution (Left and Right). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 647, ab150083) secondary antibody was used at 1/5000 dilution.

This data was developed using ab284040, the same antibody clone in a different buffer formulation.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] - BSA and Azide free (AB284054)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labelling SARS-CoV-2 orf3d with primary antibody anti-SARS-CoV-2 orf3d (ab284040) at 1/100 dilution, followed by Goat Anti-Rabbit (Alexa Fluor^® 594) (ab150089) (green) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in HEK-293T cells transfected with a SARS-CoV2 orf3d expression vector containing a myc tag, no staining in HEK-293T cells transfected with a SARS-CoV2 orf8 expression vector containing a myc tag. Myc-Tag Mouse mab (Alexa Fluor^® 647) (red) was used to counterstain cells at 1/100 dilution. The nuclear counter stain is DAPI (blue).

This data was developed using ab284040, the same antibody clone in a different buffer formulation.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] - BSA and Azide free (AB284054)

This data was developed using ab284040, the same antibody clone in a different buffer formulation.

Indirect ELISA showing primary antibody ab284040 binding to SARS-CoV2 ORF3d peptide (aa 12-32) . Plates were coated with SARS-CoV2 ORF3d peptide (aa 12-32) and recombinant SARS-CoV2 ORF8 (aa 16-121) protein (His tag) at 1000 ng/ml. Binding of ab284040 was assessed in a serial dilution range 0.016- 1000 ng/mL (a 3-fold serial dilution).

Binding was detected using pre-adsorbed secondary antibody, goat anti-rabbit IgG H&L (HRP, ab97080) at 1/2000 dilution.

• WB

Supplier Data

Western blot - Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] - BSA and Azide free (AB284054)

This data was developed using ab284040, the same antibody clone in a different buffer formulation.

ab284040 was shown to bind specifically to SARS-CoV2 Orf3d peptide in Western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Blocking buffer : 5% BSA in TBS-0.1% Tween^® 20 (TBS-T)

All lanes:

Western blot - Anti-SARS-CoV-2 orf3d antibody [EPR25045-51] (<a href='/en-us/products/primary-antibodies/sars-cov-2-orf3d-antibody-epr25045-51-ab284040'>ab284040</a>) at 1/1000 dilution

Lane 1:

Orf3d peptide (aa12-32) at 0.5 µg

Lane 2:

Orf3d peptide (aa12-32) at 0.2 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/20000 dilution

Observed band size: 6 kDa

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