Anti-GLTSCR1 抗体 [EPR26561-76] (BSA and Azide free) (ab302713)

All lanes : Anti-GLTSCR1 antibody [EPR26561-76] (ab302712) at 1/1000 dilution

Lane 1 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 2 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Performed under non-reducing conditions.

Predicted band size: 158 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?


Exposure time: 114 seconds

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

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

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. 

The membrane was incubateded with primary antibodies overnight at 4 °C
Low expression: U-87 MG (PMID: 29374058). 

The expression profile observed is consistent with what has been described in the literature (PMID: 29374058).

All lanes : Anti-GLTSCR1 antibody [EPR26561-76] (ab302712) at 1/1000 dilution

Lane 1 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Performed under non-reducing conditions.

Predicted band size: 158 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

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

Lane 2 and 3 of this blot were developed using a high sensitivity ECL substrate.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling GLTSCR1 with AB302712 at 1/100 dilution (4.91 μg/ml), followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human pancreas was observed. The section was incubated with AB302712 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling GLTSCR1 with AB302712 at 1/100 dilution (4.91 μg/ml), followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human placenta was observed. The section was incubated with AB302712 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling GLTSCR1 with AB302712 at 1/500 dilution (0.982 μg/ml), followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse pancreas was observed. The section was incubated with AB302712 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling GLTSCR1 with AB302712 at 1/500 dilution (0.982 μg/ml), followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebrum was observed. The section was incubated with AB302712 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of paraffin-embedded mouse head and neck squamous cell carcinoma tissue labeling GLTSCR1  with AB302712 at 1/500 dilution (0.982 μg/ml), followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse head and neck squamous cell carcinoma was observed. The section was incubated with AB302712 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labeling GLTSCR1 with AB302712 at 1/ 100 dilution (4.91 μg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing nuclear staining in HEK-293 cell line. Low expression: U-87 MG (PMID: 29374058) showed no staining. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/mL)(Red). The Nuclear counterstain was (Blue). 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling GLTSCR1 with AB302712 at 1/ 100 dilution (4.91 μg/mL), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/mL)(Red). The Nuclear counterstain was (Blue). 

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell, Left) / HEK-293 (human embryonic kidney epithelial cell, Right) cells labeling GLTSCR1 with ab302712 at 1/500 dilution (0.1 µg) (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). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression: U-87 MG (PMID: 29374058) was evaluated. 

This data was developed using ab302712, the same antibody clone in a different buffer formulation. 

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling GLTSCR1 with ab302712 at 1/500 dilution (0.1 µg) (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). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.