Anti-HSV1 gH antibody [BBH1]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSV1 gH antibody [BBH1] (AB110227)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue staning HSV1 gH with ab110227 at 1/1000 dilution (0.894 µg/ml), followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

Negative control : no staining on human cerebrum.

The section was incubated with ab110227 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSV1 gH antibody [BBH1] (AB110227)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a Herpes simplex virus 1 gH expression vector containing a myc-his tag. (B) HEK-293T cells transfected with empty vector containing a myc-his tag staning HSV1 gH with ab110227 at 1/1000 dilution (0.894 µg/ml), followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

Positive staining on (A) HEK-293T transfected with a Herpes simplex virus 1 gH expression vector containing a myc-his tag. No staining on (B) HEK-293T cells transfected with empty vector containing a myc-his tag.

The section was incubated with ab110227 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HSV1 gH antibody [BBH1] (AB110227)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) transfected with myc-tagged Herpes simplex virus 1 gH expression vector cells labelling HSV1 gH with ab110227 at 1/200 (4.47 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic and membranous staining in 293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Anti-Myc Rabbit polyclonal antibody (ab9106) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

• IP

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Immunoprecipitation - Anti-HSV1 gH antibody [BBH1] (AB110227)

HSV1 gH was immunoprecipitated from 0.35 mg of 293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His- tag whole cell lysate with ab110227 at 1/50 dilution (2μg in 0.35mg lysates). Western blot was performed from the immunoprecipitate using ab110227 at 1/1000 dilution. ab131366, VeriBlot for IP secondary antibody(HRP) was used as secondary antibody at 1/5000 dilution.

Lane 1(Input) : 293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag whole cell lysate, 10 μg

Lane 2(+) : 293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag whole cell lysate

Lane 3(-) : Mouse IgG2a kppa monoclonal isotype control (ab18413) instead of ab110227 in 293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag whole cell lysate

Observed MW(KDa) : 100

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

All lanes:

Immunoprecipitation - Anti-HSV1 gH antibody [BBH1] (ab110227) at 1/50 dilution

All lanes:

293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 100 kDa

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Exposure time: 180s

• WB

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Western blot - Anti-HSV1 gH antibody [BBH1] (AB110227)

Western blot analysis of ab110227.

Each track was loaded with 2x10^5 cells infected with HSV1 strain SC16, or mock-infected cells.

BBH1 - ab110227, anti-gH monoclonal antibody (1mg/ml)
Shabba - anti-gH polyclonal antibody
LP14 - anti-gD polyclonal antibody

All lanes:

Western blot - Anti-HSV1 gH antibody [BBH1] (ab110227)

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• WB

Supplier Data

Western blot - Anti-HSV1 gH antibody [BBH1] (AB110227)

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

ab213204 was used as a total protein control at 1/5000 dilution.

ab181602 was used as a GAPDH loading control at 1/200000 dilution.

Cross-reactivity has not been checked and this antibody might cross with HSV-2 gH.

All lanes:

Western blot - Anti-HSV1 gH antibody [BBH1] (ab110227) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Herpes simplex virus 1 gH expression vector containing a myc-His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 100 kDa,36 kDa

false

Exposure time: 48s