Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric)

• I-ELISA

Lab

Indirect ELISA - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (AB322999)

Indirect ELISA showing primary antibody ab322999 binding to Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Fc Chimera, ab272105). Plates were coated with Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Fc Chimera, ab272105), Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S2 (Fc Chimera, ab272106) and Recombinant Human coronavirus SARS spike glycoprotein (Tagged, ab270844) at 1000 ng/ml. Binding of ab322999 was assessed in a serial dilution range 0.016- 1000 ng/mL (a 3-fold serial dilution). Secondary antibody, Goat Anti-Chicken IgY H&L (HRP, ab6877) was used at 1 : 5000.

• WB

Lab

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (AB322999)

Lane 1 : Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active, His tagged) (ab273068) at 0.2 ug

Lane 2 : SARS-CoV Spike S1+S2 ECD-His Recombinant Protein at 0.2 ug

Performed under reducing conditions.

Observed band size : 120-160 kDa.

False colour image of Western blot : Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (ab322999) staining at 1/500 dilution, shown in black; Mouse Monoclonal 6X His tag® (ab18184) loading control staining at 1/1000 dilution, shown in red. In Western blot, ab322999 was shown to bind specifically to SARS-CoV and SARS-CoV-2 Spike Glycoproteins. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk 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. Secondary antibodies used were Goat Anti-Chicken IgY H&L (HRP) preadsorbed (ab7118) at 1 : 10000 and Goat anti-Mouse IgG H&L 680RD at 1 : 20000.

Blocking buffer : 3% milk in TBS-0.1% Tween® 20 (TBS-T).

Exposure time : 2 min.

All lanes:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - Chicken IgY (Chimeric) (ab322999) at 1/500 dilution

Lane 1:

Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (<a href='/en-us/products/proteins-peptides/recombinant-human-coronavirus-sars-cov-2-spike-glycoprotein-s1-active-ab273068'>ab273068</a>) at 0.2 µg

Lane 2:

SARS-CoV Spike S1+S2 ECD-His Recombinant Protein at 0.2 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Chicken IgY H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-preadsorbed-ab7118'>ab7118</a>) at 1/10000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 120-160 kDa

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Exposure time: 2min