Anti-SARS-CoV-2 M protein antibody [8]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 M protein antibody [8] (AB325776)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling SARS2-CoV2 Membrane Protein M with ab325776 at 1/500 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a SARS-CoV2 membrane (M) protein and SARS-CoV membrane (M) protein expression vectors containing a myc-His-tag® (shown in green), showing no staining in 293T cells transfected with a MERS-Cov membrane (M) protein and HCoV-OC43 membrane (M) protein, HCoV-HKU1 membrane (M) protein expression vectors containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Panel A : 293T cells transfected with an empty expression vector containing a myc-His-tag®.
Panel B : 293T cells transfected with a SARS-CoV2 membrane (M) protein expression vector containing a myc-His-tag®.
Panel C : 293T cells transfected with a SARS-CoV membrane (M) protein expression vector containing a myc-His-tag®.
Panel D : 293T cells transfected with a MERS-Cov membrane (M) protein expression vector containing a myc-His-tag®.
Panel E : 293T cells transfected with a HCoV-OC43 membrane (M) protein expression vector containing a myc-His-tag®.
Panel F : 293T cells transfected with a HCoV-HKU1 membrane (M) protein expression vector containing a myc-His-tag®.

ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• I-ELISA

Lab

Indirect ELISA - Anti-SARS-CoV-2 M protein antibody [8] (AB325776)

Indirect ELISA analysis of ab325776 at 2000 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 1000 dilution dilution.

Antigen A : 293T cells (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®.
Antigen B : 293T transfected with a SARS-CoV2 membrane (M) protein expression vector containing a myc-His-tag®.
Antigen C : 293T transfected with a SARS-CoV membrane (M) protein expression vector containing a myc-His-tag®.
Antigen D : 293T transfected with a MERS-Cov membrane (M) protein expression vector containing a myc-His-tag®.
Antigen E : 293T transfected with a HCoV-OC43 membrane (M) protein expression vector containing a myc-His-tag®.
Antigen F : 293T transfected with a HCoV-HKU1 membrane (M) protein expression vector containing a myc-His-tag®.

Antigen concentration : 1000-0 ug/ml

• WB

Lab

Western blot - Anti-SARS-CoV-2 M protein antibody [8] (AB325776)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

In Western blot,Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (25kDa).

All lanes:

Western blot - Anti-SARS-CoV-2 M protein antibody [8] (ab325776) at 1/5000 dilution

Lane 1:

293T cells (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with a SARS-CoV2 membrane (M) protein expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

293T transfected with a SARS-CoV membrane (M) protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 4:

293T transfected with a MERS-Cov membrane (M) protein expression vector containing a myc-His-tag®, whole cell lysate at 4 µg

Lane 5:

293T transfected with a HCoV-OC43 membrane (M) protein expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 6:

293T transfected with a HCoV-HKU1 membrane (M) protein expression vector containing a myc-His-tag®, whole cell 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 1/20000 dilution

Observed band size: 25 kDa,36 kDa

true

Exposure time: 37s