Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling GBA(mutated E326K) with ab302607 at 1/5000 (0.094 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : No staining on human breast. The section was incubated with ab302607 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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded A HEK-293T cells tr tissue labeling GBA(mutated E326K) with ab302607 at 1/5000 (0.094 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on HEK-293T cells transfected with a human GBA E365K expression vector containing a myc-His tag cell pallet (image A). No staining on HEK-293T cells transfected with a human WT GBA expression vector containing a myc-His tag cell pallet (image B) and 293T transfected with an empty vector cell pallet (image C). The section was incubated with ab302607 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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labelling GBA(mutated E326K) with ab302607 at 1/1000 (0.472 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 cytoplasmic staining in HEK-293T cells transfected with a human GBA E365K expression vector containing a Myc tag and no staining in HEK-293T cells transfected with a human GBA expression vector containing a Myc tag is observed. 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 2 ug/mL dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) cells transfected with a human GBA (WT) expression vector containing a myc-his tag (Left) / HEK-293T transfected with a human GBA (mutated E326K) expression vector containing a myc-his tag (Right) cells labelling GBA(mutated E326K) with ab302607 at 1/500 dilution (0.1ug)/ Left and Right (Red) compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative in GBA (WT) transfected cells and GBA (WT)-expressing tissues. Exposure time : 3 minutes

All lanes:

Western blot - Anti-GBA (mutated E326K) antibody [EPR24900-273] (<a href='/en-us/products/primary-antibodies/gba-mutated-e326k-antibody-epr24900-273-ab302607'>ab302607</a>) at 1/1000 dilution

Lane 1:

HEK-293T transfected with a human GBA (E326K mutation) expression vector containi a His-tag, whole cell lysate 20 µg

Lane 2:

HEK-293T transfected with a human GBA (WT) expression vector containi a His-tag, whole cell lysate 20 µg

Lane 3:

Human spleen tissue lysate 20 µg

Lane 4:

Human fetal brain tissue lysate 20 µg

Lane 5:

Human adrenal gland tissue lysate 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 1/100000 dilution

Observed band size: 60 kDa

false

Exposure time: 3min

• Dot

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Dot Blot - Anti-GBA (mutated E326K) antibody [EPR24900-273] (BSA and Azide free) (AB302608)

This data was developed using ab302607, the same antibody clone in a different buffer formulation. Dot blot analysis of GBA(mutated E326K) using ab302607 at 1 : 1000 (0.472 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 3 minutes Blocking and diluting buffer and concentration : 5% NFDM/TBST