Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (AB322461)

Immunohistochemical analysis of paraffin-embedded (A) HEK-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®. (B) HEK-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®. (C) HEK-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®. (D) HEK-293T transfected with an empty vector containing a His-tag®. tissue labeling HBV envelope L protein with ab322461 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) HEK-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®. No staining in (B) HEK-293T transfected with Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Middle (M) envelope protein expression vector containing a myc-His-tag®, (C) HEK-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 (D) HEK-293T transfected with an empty vector containing a His-tag.
The section was incubated with ab322461 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (AB322461)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling HBV envelope L protein with ab322461 at 1/50 (10.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in 293T cells (shown in green) transfected with a Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Large (L) envelope protein expression vector containing a myc-His-tag®. 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).

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

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.

• I-ELISA

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Indirect ELISA - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (AB322461)

Indirect ELISA analysis of ab322461 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Large envelope protein.

Antigen concentration : 1000 ng/ml

• WB

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Western blot - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (AB322461)

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

This antibody reacts with Hepatitis B virus Large (L) envelope protein from genotype A2 but not genotype H, D or A1.

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (ab322461) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a Hepatitis B virus genotype A2 Large (L) envelope protein expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

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 20 µg

Lane 4:

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 20 µg

Lane 5:

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 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 44 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (AB322461)

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

This antibody does not cross-react with overexpressed Hepatitis B virus genotype A2 subtype adw2 (strain Rutter 1979) Middle (M) envelope protein and Small (S) envelope protein.

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Hepatitis B Virus Surface Antigen antibody [EPR29887-532] (ab322461) at 1/1000 dilution

Lane 1:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 44 kDa,36 kDa

false

Exposure time: 180s