Anti-GILT antibody [EPR30714-572]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) IMR-32 (human neuroblastoma neuroblast) cells labelling GILT with ab325920 at 1/500 (0.968 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 staining in Raji cell line (green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Low expression : IMR-32.

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325920 at 1/500 dilution, followed by ab150120 at 1/1000 dilution.-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling GILT with ab325920 at 1/5000 (0.097 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal cells of human liver cancer.

The primary antibody was incubated for 10 mins at room temperature, Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling GILT with ab325920 at 1/5000 (0.097 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast cancer (PMID : 34648659).

The primary antibody was incubated for 10 mins at room temperature, Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunohistochemical analysis of paraffin-embedded (A) : Raji (human Burkitt's lymphoma B lymphocyte) cell pellet (B) : IMR-32 (human neuroblastoma neuroblast) cell pellet tissue labeling GILT with ab325920 at 1/1000 (0.484 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Raji cell pellet, no staining on IMR-32 cell pellet.

The primary antibody was incubated for 10 mins at room temperature, Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling GILT with ab325920 at 1/5000 (0.097 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil (PMID : 24409177).

The primary antibody was incubated for 10 mins at room temperature, Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

• WB

Lab

Western blot - Anti-GILT antibody [EPR30714-572] (AB325920)

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

Low expression : skeletal muscle.

The weak band observed at ~35KDa may represent the precursor form of GILT (PMID : 39273610).

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

All lanes:

Western blot - Anti-GILT antibody [EPR30714-572] (ab325920) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human breast cancer tissue lysate at 20 µg

Lane 3:

Human lung tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Lane 5:

Human lymph node tissue lysate at 20 µg

Lane 6:

Human tonsil tissue lysate at 50 µg

Lane 7:

Human lung tissue lysate at 50 µg

Lane 8:

Human skeletal muscle tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 28 kDa,35 kDa,36 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-GILT antibody [EPR30714-572] (AB325920)

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

Low expression : IMR-32, SK-N-BE(2).

The weak band observed at ~35KDa may represent the precursor form of GILT (PMID : 39273610).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-GILT antibody [EPR30714-572] (ab325920) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

SK-N-BE(2) (human neuroblastoma neuroblast) 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 1/20000 dilution

Observed band size: 28 kDa,35 kDa,124 kDa

false

Exposure time: 3s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GILT antibody [EPR30714-572] (AB325920)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling GILT with ab325920 at 1/5000 (0.097 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle.

The primary antibody was incubated for 10 mins at room temperature, Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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