Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling AAV2 Capsid protein VP1 with ab315287 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human cerebrum.

The section was incubated with ab315287 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling AAV2 Capsid protein VP1 with ab315287 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cerebrum.

The section was incubated with ab315287 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling AAV2 Capsid protein VP1 with ab315287 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat cerebrum.

The section was incubated with ab315287 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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with a VVA2-VP1 expression vector containing a myc tag cells labelling AAV2 Capsid protein VP1 with ab315287 at 1/50 (10.6 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 nuclear staining in 293T transfected with a VVA2-VP1 expression vector containing a myc tag.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38ug/ml dilution (Red). 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A)HEK-293T (human embryonic kidney epithelial cell) transfected with a VVA2-VP1 expression vector containing a myc tag. (B) HEK-293T transfected with an empty vector. tissue labeling AAV2 Capsid protein VP1 with ab315287 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a VVA2-VP1 expression vector containing a myc tag, no staining on (B) HEK-293T transfected with an empty vector.

The section was incubated with ab315287 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T transfected with a VVA2-VP1 expression vector containing a myc tag (Middle) / 293T transfected with an empty expression vector containing a myc tag (Right) cells labelling AAV2 Capsid protein VP1 with ab315287 at 1/50 dilution (1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

AAV2 Capsid protein VP1 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with a AAV-2 VP1 expression vector containing a myc-His-tag®, whole cell lysate with ab315287 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315287 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) cells transfected with a AAV-2 VP1 expression vector containing a myc-His-tag®, whole cell lysate

Lane 2 : ab315287 IP in 293T (human embryonic kidney epithelial cell) cells transfected with a AAV-2 VP1 expression vector containing a myc-His-tag®, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315287 in 293T cells transfected with a AAV-2 VP1 expression vector containing a myc-His-tag®, whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] (<a href='/en-us/products/primary-antibodies/aav2-capsid-protein-vp1-antibody-epr28595-59-ab315287'>ab315287</a>) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) cells transfected with a AAV-2 VP1 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

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-AAV2 Capsid protein VP1 antibody [EPR28595-59] - BSA and Azide free (AB315288)

This data was developed using ab315287, the same antibody clone in a different buffer formulation.

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

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-AAV2 Capsid protein VP1 antibody [EPR28595-59] (<a href='/en-us/products/primary-antibodies/aav2-capsid-protein-vp1-antibody-epr28595-59-ab315287'>ab315287</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containi a myc-His-tag®, whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a AAV-2 VP1 expression vector containi a myc-His-tag®, whole cell lysate at 5 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: 81 kDa,36 kDa

false

Exposure time: 10s