重组Anti-SARS-CoV-2 Spike Glycoprotein S1抗体[8B12-C2] - BSA and Azide free

• ELISA

Lab

ELISA - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - BSA and Azide free (AB282749)

Plates were coated with Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Fc Chimera, ab272105) and Recombinant Human coronavirus SARS-CoV-2 Spike Glycoprotein S2 (Fc Chimera, ab272106) at 1000 ng/ml.
Primary antibody (ab282749) concentration range 0.045-100 ng/ml.
Secondary antibody, Peroxidase AffiniPure Goat Anti-Mouse IgG, Fcγ Fragment Specific used at 1/2000 dilution.

• WB

Collaborator

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - BSA and Azide free (AB282749)

Lanes 1 - 3:

anti-P2A at 1/500 dilution

Lanes 4 - 6:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - BSA and Azide free (ab282749) at 0.1 µg/mL

Lanes 1 and 4:

HEK293T whole cell lysate

Lanes 2 and 5:

Whole cell lysates of HEK293T cells expressing wild-type Sars-CoV2 Spike-P2A-GFP protein

Lanes 3 and 6:

Whole cell lysates of HEK293T cells expressing the B 1.351 variant of Sars-CoV2 Spike-P2A-GFP protein

Secondary

All lanes:

HRP-coupled

false

• WB

Supplier Data

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - BSA and Azide free (AB282749)

Lanes 1 - 5 :  Green – ab282749 at 1 in 500 dilution, observed from 120 – 180 kDa
Lanes 6 - 8 :  Red – Rabbit polyclonal to 6X His tag^® (ab9108) at 1 in 1000 dilution, observed from 120 – 160 kDa

Lanes 9 & 10 : Black – Goat polyclonal to DDDDK tag (Binds to FLAG^® tag sequence) at 1 μg/mL, observed at 180 kDa

ab282749 was shown to bind specifically to SARS-CoV and SARS-CoV-2 Spike Glycoproteins 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.

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

Lanes 1 - 5:

Western blot - Anti-SARS-CoV-2 Spike Glycoprotein S1 antibody [8B12-C2] - BSA and Azide free (ab282749) at 1/500 dilution

Lanes 6 - 8:

Western blot - Anti-6X His tag® antibody (<a href='/products/primary-antibodies/6x-his-tag-antibody-ab9108'>ab9108</a>) at 1/1000 dilution

Lanes 9 - 10:

Goat polyclonal to DDDDK tag at 1 µg/mL

Lanes 1 and 6:

Western blot - Recombinant human coronavirus SARS-CoV-2 Spike Glycoprotein S1 (Active) (<a href='/products/proteins-peptides/recombinant-human-coronavirus-sars-cov-2-spike-glycoprotein-s1-active-ab273068'>ab273068</a>) at 0.5 µg

Lanes 2 and 7:

SARS-CoV-2 (2019-nCoV) Spike S1+S2 ECD-His Recombinant Protein at 0.2 µg

Lanes 3 and 8:

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

Lanes 4 and 9:

SARS-CoV-2 3xFlag Spike Protein Transfected Expi whole cell lysate at 20 µg

Lanes 5 and 10:

SARS-CoV 3xFlag Spike Protein Transfected Expi whole cell lysate at 20 µg

Secondary

Lanes 1 - 5:

Western blot - Donkey anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/donkey-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216778'>ab216778</a>) at 1/20000 dilution

Lanes 6 - 8:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/20000 dilution

Lanes 9 - 10:

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

Observed band size: 120-180 kDa,120-160 kDa,180 kDa

false