Anti-Glucose Oxidase antibody [EPR27335-25]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a GOX expression vector containing a myc tag (Middle) / 293T cells transfected with a empty expression vector containing a myc tag (Right) cells labelling Glucose Oxidase with ab314545 at 1/5000 dilution (0.01 ug)/Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidneyepithelial cell) transfected with a GOX expression vector containing a myc tag cells labelling Glucose Oxidase with ab314545 at 1/1000 (0.507 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1008 (2ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with a GOX 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 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 1/1008 (2ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Immunohistochemical analysis of paraffin-embedded Panel A. HEK-293T cells transfected with a GOX expression vector containing a myc-His tag. Panel B. HEK-293T cells transfected with an empty vector. tissue labeling Glucose Oxidase with ab314545 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on HEK-293T cells transfected with a GOX expression vector containing a myc-His tag (Panel A). No staining on HEK-293T cells transfected with an empty vector (Panel B). The section was incubated with ab314545 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Glucose Oxidase with ab314545 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human cerebrum. The section was incubated with ab314545 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.

• I-ELISA

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Indirect ELISA - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Indirect ELISA analysis of ab314545 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 : Aspergillus niger Glucose Oxidase. Antigen concentration : 1000 ng/ml

• IP

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Immunoprecipitation - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

Glucose Oxidase was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with an Aspergillus niger gox expression vector containing a His-tag, whole cell lysate with ab314545 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314545 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 an Aspergillus niger gox expression vector containing a His-tag, whole cell lysate Lane 2 : ab314545 IP in 293T (human embryonic kidney epithelial cell) cells transfected with an Aspergillus niger gox expression vector containing a His-tag, whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314545 in 293T cells transfected with an Aspergillus niger gox expression vector containing a His-tag, whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

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Immunoprecipitation - Anti-Glucose Oxidase antibody [EPR27335-25] (ab314545) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) cells transfected with an Aspergillus niger gox expression vector containing a His-tag, whole cell lysate

Secondary

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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: 5s

• WB

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Western blot - Anti-Glucose Oxidase antibody [EPR27335-25] (AB314545)

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.

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Western blot - Anti-Glucose Oxidase antibody [EPR27335-25] (ab314545) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 5 µg

Lane 2:

293T cells transfected with an Aspergillus niger gox expression vector containing a 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: 80 kDa

false

Exposure time: 10s