Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116]

• ELISA

Lab

ELISA - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Indirect ELISA showing primary antibody ab283942 binding to recombinant human coronavirus SARS-CoV-2 (Omicron variant) Spike Glycoprotein S1; SARS-CoV-2 (Omicron variant) Spike Glycoprotein RBD; SARS-CoV-2 Omicron variant Spike protein (GCN4-IZ) His tag; Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Fc Chimera).

Plates were coated with recombinant human coronavirus SARS-CoV-2 (Omicron variant) Spike Glycoprotein S1; SARS-CoV-2 (Omicron variant) Spike Glycoprotein RBD; SARS-CoV-2 Omicron variant Spike protein (GCN4-IZ) His tag; Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Fc Chimera) at 1000 ng/mL. Binding of ab283942 was assessed in a serial dilution range 0 - 1000 ng/mL.

Binding was detected usingAlkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution.

Substrate Solution : p-nitrophenyl phosphate(PNPP)

• ELISA

Supplier Data

ELISA - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Indirect ELISA showing primary antibody ab283942 binding to the antigen ab272105 (recombinant human coronavirus SARS-CoV-2 spike glycoprotein S1 (Fc Chimera)). Plates were coated with recombinant human coronavirus SARS-CoV-2 spike glycoprotein S1 (Fc Chimera, ab272105) and recombinant human coronavirus SARS spike glycoprotein (Tagged, ab270844) at 1000 ng/ml. Binding of ab283942 was assessed in a serial dilution range 0.04- 100 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Immunocytochemistry/ Immunofluorescence analysis of HEK-293T transfected with a Spike-SARS-CoV2 expression vector labeling SARS-CoV-2 Spike Glycoprotein S1 with ab283942 at 1/500 dilution followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary (green) at 1/1000 dilution. The counterstain was Anti-FLAG mouse monoclonal antibody at 1/500 dilution followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary(red) at 1/1000 dilution. Cells were fixed with 100% methanol and permeabilised with 0.1% TritonX-100. Nuclear counterstain was DAPI. Confocal image showing the antibody co-stain with Flag in HEK-293T cells transfected with a Spike-SARS-CoV2 expression vector containing a flag tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt

Lab

Flow Cytometry - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a Spike SARS-CoV-1 (Left) or Spike SARS-CoV-2 (Right) expression vector labeling SARS-CoV-2 Spike Glycoprotein S1 with ab283942 at 1/500 dilution followed by ab150081 AlexaFluor®488 Goat anti-Rabbit secondary (green) at 1/2000 dilution. Cells were fixed with 2% Paraformaldehyde and permeabilised with 0.1% Tween-20. Cells were surface stained with ab283942. Then fixed with 2% PFA for 10min followed by intracellularly stained with anti-Flag tag conjugated to BV421. No cross-reactivity with Spike SARS-CoV-1.

• WB

Supplier Data

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Lane 1 : Red – loading control Mouse anti-6x His tag antibody (ab18184) observed at 135 kDa

Lanes 2 : Green – ab283942 observed at 135 kDa

ab283942 was shown to bind specifically to SARS-CoV-2 spike glycoprotein S1 in Western blot. 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-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at 1/20000 dilution.

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

All lanes:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (ab283942) at 1/1000 dilution

All lanes:

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

Observed band size: 135 kDa

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• WB

AbReview79645****

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

All lanes:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (ab283942) at 1 µg/mL

Lane 1:

Precision Plus Protein™ Dual Color Standard

Lane 2:

CaCo2 Uninfected whole cell lysate, at 40 µg

Lane 3:

CaCo2 infected with SARS-CoV-2 whole cell lysate at 40 µg

Lane 4:

CaCo2 infected with CoV-NL63 whole cell lysate at 40 µg

Lane 5:

CaCo2 infected with CoV-OC463 whole cell lysate at 40 µg

Lane 6:

recombinant SARS CoV-2 spike protein + twin strep tag at 75 ng

Lane 7:

A549 WT whole cell lysate at 15 µg

Secondary

All lanes:

LiCor goat anti-rabbit IRDye; 800CW

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This image is courtesy of an Abreview submitted by Nicola Goodman

• WB

Supplier Data

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

ab283942 was shown to bind specifically to SARS-CoV-2 spike glycoprotein S1 in Western blot. 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 Donkey anti-Goat IgG H&L (IRDye® 800CW) preadsorbed (ab216775) and Donkey anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216779) at 1/20000 dilution.

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

Lanes 1 - 2:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (ab283942) at 1/20000 dilution

Lanes 3 - 4:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (ab283942) at 1/1000 dilution

Lanes 1 and 3:

Expi293 cells transfected with SARS-CoV2 3xFlag Spike Protein at 20 µg

Lanes 2 and 4:

Expi293 cells transfected with SARS-CoV1 3xFlag Spike Protein at 20 µg

Secondary

All lanes:

Western blot - Donkey anti-Goat IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/donkey-goat-igg-h-l-irdye-800cw-preadsorbed-ab216775'>ab216775</a>) at 1/20000 dilution

Observed band size: 200 kDa

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• Dot

Lab

Dot Blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [EPR24852-116] (AB283942)

Dot blot analysis of Recombinant Human coronavirus SARS-CoV-2 (B.1.1.529/Omicron) Spike Glycoprotein S1 (ab290828) (Lane 1), Recombinant Human coronavirus SARS-CoV-2 (B.1.1.529/Omicron) Spike Glycoprotein S1 RBD protein (ab290829) (Lane 2), Recombinant Human coronavirus SARS-CoV-2 (B.1.1.529/Omicron) Spike protein (Lane 3) and Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (ab272105) (Lane 4) labelling SARS-CoV-2 Spike Glycoprotein S1 with ab283942 at a dilution of 1/1000. Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG was used as the secondary antibody at a dilution of 1/2000. This antibody reacts with Omicron (B.1.1.529) variant. Blocking and dilution buffer : 5% NFDM/TBST. Exposure time : 180 seconds.