Anti-GBP5 antibody [EPR28367-83]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling GBP5 with ab313390 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab313390 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Human colon carcinom tissue labeling GBP5 with ab313390 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of human colon carcinoma. The section was incubated with ab313390 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling GBP5 with ab313390 at 1/4000 (0.127 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : no staining on human cerebrum (PMID : 33608513). The section was incubated with ab313390 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded (A) THP-1 (human mon tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) THP-1 treated with 1ug/ml LPS and IFN gamma for 24 hours cell pellet, no staining on (B) Untreated THP-1 cell pellet. The section was incubated with ab313390 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling GBP5 with ab313390 at 1/50 (10.14 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 THP-1 cells treated with 1 µg/mL LPS (lipopolysaccharide) and 40 ng/mL IFN gamma for 24 hours, while showing no staining in untreated THP-1 cells.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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-GBP5 antibody [EPR28367-83] (AB313390)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte) treated with 1µg/mL LPS for 24h and then 40ng/mL IFN gamma for 24h (Red) / untreated THP-1 (Green) cells labelling GBP5 with ab313390 at 1/500 dilution (0.1 ug)/Red and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) 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 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : no staining on rat cerebrum. The section was incubated with ab313390 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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)

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Flow Cytometry (Intracellular) - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) treated with 1µg/mL LPS for 24h and then 40ng/mL IFN gamma for 24h (Red) / untreated J774A.1 (Green) cells labelling GBP5 with ab313390 at 1/500 dilution (0.1 ug)/Red and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) 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 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : no staining on mouse cerebrum. The section was incubated with ab313390 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of mouse colon. The section was incubated with ab313390 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse glioblastoma. The section was incubated with ab313390 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling GBP5 with ab313390 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of rat colon. The section was incubated with ab313390 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. 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-GBP5 antibody [EPR28367-83] (AB313390)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cells labelling GBP5 with ab313390 at 1/50 (10.14 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 J774A.1 cells treated with 1 µg/mL LPS (lipopolysaccharide) and 200 ng/mL IFN gamma (mouse) for 24 hours, while showing no staining in untreated J774A.1 cells.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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.

• WB

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Western blot - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The cell induction condition was referenced from literature(PMID : 22461501). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane1-2 : 180 seconds, lane3-4 : 15 seconds

All lanes:

Western blot - Anti-GBP5 antibody [EPR28367-83] (ab313390) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with /ml IFN alpha for 16 hours whole cell lysate at 20 µg

Lane 3:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 4:

THP-1 treated with /ml LPS and IFN gamma for 24 hours whole cell lysate at 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: 66 kDa

false

• WB

Supplier Data

Western blot - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : brain(PMID : 33608513). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-GBP5 antibody [EPR28367-83] (ab313390) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human brain tissue lysate at 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: 66 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GBP5 antibody [EPR28367-83] (AB313390)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The cell induction condition was referenced from literature(PMID : 22461501). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-GBP5 antibody [EPR28367-83] (ab313390) at 1/1000 dilution

Lane 1:

Untreated J774A.1 (Mouse reticum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 2:

J774A.1 treated with /ml LPS and IFN gamma for 24 hours, was then added for an additional 300ng/ml BFA for last 20 hours whole cell lysate at 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: 66 kDa

false

Exposure time: 15s