Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling GGT1/GGT + GGT3 with ab316768 at 1/50 (9.98 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 mainly membranous staining in HepG2 cells.
Low expression : HEK-293 (PMID : 23682772).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cardiac muscle.
The section was incubated with ab316768 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-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on bile canaliculi of human liver (PMID : 8813074).
The section was incubated with ab316768 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-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling GGT1/GGT + GGT3 with ab316768 at 1/100 (4.99 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on proximal tubules of human kidney (PMID : 8813074, 32474759).
The section was incubated with ab316768 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.

• IP

Supplier Data

Immunoprecipitation - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

GGT1/GGT + GGT3 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab316768 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316768 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : ab316768 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316768 in HepG2 whole cell lysate

All lanes:

Immunoprecipitation - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (<a href='/ko/products/primary-antibodies/ggt1-ggt-ggt3-antibody-epr28715-82-ab316768'>ab316768</a>) at 1/30 dilution

All lanes:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 76s

• WB

Supplier Data

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Amongst human GGT homologous proteins, this antibody reacts with GGT1 and GGT3 but not GGT2.

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-GGT1/GGT + GGT3 antibody [EPR28715-82] (<a href='/ko/products/primary-antibodies/ggt1-ggt-ggt3-antibody-epr28715-82-ab316768'>ab316768</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate 7 at 7 µg

Lane 2:

293T cells transfected with a human GGT1 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Lane 3:

293T cells transfected with a human GGT2 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Lane 4:

293T cells transfected with a human GGT3 expression vector containing a myc-His-tag®, whole cell lysate at 7 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60-75 kDa,36 kDa,60 kDa,75 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Low expression : skeletal muscle (PMID : 7689219).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (<a href='/ko/products/primary-antibodies/ggt1-ggt-ggt3-antibody-epr28715-82-ab316768'>ab316768</a>) at 1/1000 dilution

Lane 1:

Human kidney tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60-75 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] - BSA and Azide free (AB316769)

This data was developed using ab316768, the same antibody clone in a different buffer formulation.

Low expression : HEK-293 (PMID : 23682772).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GGT1/GGT + GGT3 antibody [EPR28715-82] (<a href='/ko/products/primary-antibodies/ggt1-ggt-ggt3-antibody-epr28715-82-ab316768'>ab316768</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

Untreated HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HepG2 whole cell lysate treated with Protein Deglycosylation Mix II, at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60-75 kDa,36 kDa

false

Exposure time: 81s