Anti-Glucose Transporter GLUT3 antibody [EPR30042-14]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) and Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labelling Glucose Transporter GLUT3 with ab327003 at 1/500 (1.006 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membranous and cytoplasmic staining in Neuro-2a cells and no staining in Beta-TC-6 cells(shown in green). The counterstain was observed in magenta.

Low expression : Beta-TC-6.

Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Magenta).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on mouse kidney.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast, Right) and Beta-TC-6 (mouse pancreas insulinoma beta cell, Left) cells labelling Glucose Transporter GLUT3 with ab327003 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

Low expression : Beta-TC-6.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on rat kidney.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Mouse lung adenocarcinoma tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung adenocarcinoma.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Glucose Transporter GLUT3 with ab327003 at 1/2000 (0.252 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis (PMID : 26299481).

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Lab

Western blot - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

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

Low expression : kidney(PMID : 1730609).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 1730609; PMID : 33061855; PMID : 9253355).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (ab327003) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat cerebellum tissue lysate at 20 µg

Lane 5:

Rat testis tissue lysate at 20 µg

Lane 6:

Rat kidney 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 1/20000 dilution

Observed band size: 49-75 kDa,36 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

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

Low expression : NIH/3T3, Beta-TC-6.

Samples are non-boiled as boiling may cause protein aggregation.

The identity of the higher MW band observed in lanes 4 is most likely glycosylated forms of Glucose Transporter GLUT3 (PMID : 10336639).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

false

Exposure time: 8s

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse testis tissue staining Glucose Transporter GLUT3 with ab327003 at a 1/1000 dilution (0.49 μg/ml), ab267377 anti-Wilms tumor protein used at a 1/500 dilution (0.766 μg/ml) and ab324729 anti-SCP1 used at a 1/2000 dilution (0.252 μg/ml).

Panel A : merged staining of anti-Glucose Transporter GLUT3 (green; Opal^™570), anti-Wilms tumor protein (magenta; Opal^™690) and anti-SCP1 (gray; Opal^™520) on mouse testis.

Panel B : anti-Glucose Transporter GLUT3 staining membrane of spermatogonia in mouse testis.

Panel C : anti-Wilms tumor protein staining nucleus of Sertoli cells in mouse testis.

Panel D : anti-SCP1 staining nucleus of primary spermatocytes in mouse testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab327003, ab267377 and ab324729 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit using Opal Polymer HRP Ms + Rb secondary reagent. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Glucose Transporter GLUT3 antibody [EPR30042-14] (AB327003)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat testis tissue staining Glucose Transporter GLUT3 with ab327003 at a 1/1000 dilution (0.49 μg/ml), ab267377 anti-Wilms tumor protein used at a 1/500 dilution (0.766 μg/ml) and ab324729 anti-SCP1 used at a 1/2000 dilution (0.252 μg/ml).

Panel A : merged staining of anti-Glucose Transporter GLUT3 (green; Opal^™570), anti-Wilms tumor protein (magenta; Opal^™690) and anti-SCP1 (gray; Opal^™520) on rat testis.

Panel B : anti-Glucose Transporter GLUT3 staining membrane of spermatogonia in rat testis.

Panel C : anti-Wilms tumor protein staining nucleus of Sertoli cells in rat testis.

Panel D : anti-SCP1 staining nucleus of primary spermatocytes in rat testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab327003, ab267377 and ab324729 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit using Opal Polymer HRP Ms + Rb secondary reagent. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.