Anti-MAG/GMA antibody [EPR24276-125]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of paraffin-embedded human sciatic nerve tissue labeling MAG/GMA with ab277524 at 1/5000 dilution (0.1008 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human sciatic nerve. The section was incubated with ab277524 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval 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-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling MAG/GMA with ab277524 at 1/5000 dilution (0.1008 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum.The section was incubated with ab277524 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

MAG/GMA was immunoprecipitated from 0.35 mg human cerebellum tissue lysate with ab277524 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277524 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human cerebellum tissue lysate 10 ug

Lane 2 : ab277524 IP in Human cerebellum tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab277524 in Human cerebellum tissue lysate

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

Exposure time : 23 seconds.

All lanes:

Immunoprecipitation - Anti-MAG/GMA antibody [EPR24276-125] (ab277524)

Predicted band size: 69 kDa

Observed band size: 100 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling MAG/GMA with ab277524 at 1/2000 dilution (0.252 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse hippocampus. (PMID : 25523827)The section was incubated with ab277524 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum tissue labeling MAG/GMA with ab277524 at 1/500 dilution (1.008 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cell cells labelling MAG/GMA with ab277524 at 1/50 dilution (10.08 ug/ml), followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in rat primary oligodendroglia cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Anti-Myelin Basic Protein rat monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab277241 at 1/50 dilution, followed by ab150157 at 1/1000 dilution.

-ve control 2 : Anti-Myelin Basic Protein rat monoclonal antibody, followed by ab150080 at 1/1000 dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver tissue labeling MAG/GMA with ab277524 at 1/100 dilution (5.04 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Negative control : No staining on rat liver (PMID2432614) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling MAG/GMA with ab277524 at 1/100 dilution (5.04 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Negative control : No staining on mouse liver (PMID2432614) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling MAG/GMA with ab277524 at 1/2000 (0.252 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat hippocampus. (PMID : 17705198, 27000654 )The section was incubated with ab277524 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling MAG/GMA with ab277524 at 1/500 dilution (1.008 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IP

Supplier Data

Immunoprecipitation - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

MAG/GMA was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab277524 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277524 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : ab277524 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab277524 in Mouse brain tissue lysate

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

Exposure time : 23 seconds.

All lanes:

Immunoprecipitation - Anti-MAG/GMA antibody [EPR24276-125] (ab277524)

Predicted band size: 69 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

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

MAG is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 1716323).

Negative control : Liver (PMID : 2432614).

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-MAG/GMA antibody [EPR24276-125] (ab277524) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 69 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-MAG/GMA antibody [EPR24276-125] (AB277524)

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

MAG is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 1716323).

Negative control : Liver (PMID : 2432614).

Exposure time : 126 seconds.

All lanes:

Western blot - Anti-MAG/GMA antibody [EPR24276-125] (ab277524) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Rat cerebellum tissue lysate at 20 µg

Lane 5:

Rat liver 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

Predicted band size: 69 kDa

Observed band size: 100 kDa

false