Anti-HBV envelope L+M+S protein antibody [EPR29889-558]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (AB324674)

Immunohistochemical analysis of paraffin-embedded Panel A 293T (human embryonic kidney epithelial cell) transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) envelope protein expression vector containing a myc-His-tag®, Panel B 293T transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Middle (M) envelope protein expression vector containing a myc-His-tag®, Panel C 293T transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Small (S) envelope protein expression vector containing a myc-His-tag® and Panel D 293T cells transfected with an empty vector containing a Myc-his tag® labeling Hepatitis B Virus Surface Antigen with ab324674 at 1/5000 (0.092 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in 293T transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) envelope protein expression vector containing a myc-His-tag® (Panel A); Positive staining in 293T transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Middle (M) envelope protein expression vector containing a myc-His-tag® (Panel B); Positive staining in 293T transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Small (S) envelope protein expression vector containing a myc-His-tag® (Panel C); No staining in 293T transfected with an empty vector containing a myc-His-tag® (Panel D).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (AB324674)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Hepatitis B Virus Surface Antigen with ab324674 at 1/2000 (0.229 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing positive staining in 293T cells (shown in green) transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) protein, Medium (M) protein and Small (S) protein expression vector containing a myc-His-tag® respectively. The counterstain was observed in magenta. Nuclear DNA was labelled 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 Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) protein expression vector containing a myc-His-tag®.
Panel C : 293T cells transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Medium (M) protein expression vector containing a myc-His-tag®.
Panel D : 293T cells transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Small (S) protein expression vector containing a myc-His-tag®.

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 (5 ug/ml) dilution (Magenta).

• I-ELISA

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Indirect ELISA - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (AB324674)

Indirect ELISA analysis of ab324674 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : HBV envelope S protein.

Antigen concentration : 1000 ng/ml

• WB

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Western blot - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (AB324674)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (ab324674) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a Myc-his tag®, whole cell lysate at 2 µg

Lane 2:

293T cells transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 3:

293T cells transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Middle (M) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 4:

293T cells transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Small (S) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 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 1/100000 dilution

Observed band size: 25 kDa,36 kDa,15-50 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (AB324674)

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

This antibody cross-reacts with overexpressed Hepatitis B virus genotype A1 Large (L) envelope protein.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HBV envelope L+M+S protein antibody [EPR29889-558] (ab324674) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 2:

293T cells transfected with a Hepatitis B virus genotype H Large (L) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 3:

293T cells transfected with a Hepatitis B virus genotype D Large (L) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 µg

Lane 4:

293T cells transfected with a Hepatitis B virus genotype A1 Large (L) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 2 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 1/100000 dilution

Observed band size: 45 kDa,36 kDa,44 kDa,31 kDa

false

Exposure time: 3s