Recombinant Anti-MAG/GMA antibody [EPR24276-125] (ab277524)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24276-125] to MAG/GMA
- Suitable for: WB, IHC-P, IHC-Fr, IP, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-MAG/GMA antibody [EPR24276-125]
See all MAG/GMA primary antibodies -
Description
Rabbit monoclonal [EPR24276-125] to MAG/GMA -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IHC-Fr, IP, ICC/IFmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human cerebellum tissue lysate; Mouse brain and cerebellum tissue lysates; Rat cerebellum tissue lysate. IHC-P: Human cerebellum and sciatic nerve tissue; Mouse hippocampus tissue; Rat hippocampus tissue. IHC-Fr: Mouse cerebellum tissue; Rat cerebellum tissue. ICC/IF: Rat primary neural/glia cells. IP: Human cerebellum tissue lysate; Mouse brain tissue lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24276-125 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab277524 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 69 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Can be used at 1/5000 dilution for human. |
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IHC-Fr |
1/500.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
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IP |
1/30.
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ICC/IF |
1/50.
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Notes |
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WB
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 69 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Can be used at 1/5000 dilution for human. |
IHC-Fr
1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
IP
1/30. |
ICC/IF
1/50. |
Target
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Function
Adhesion molecule in postnatal neural development that mediates sialic-acid dependent cell-cell interactions between neuronal and myelinating cells. Preferentially binds to alpha-2,3-linked sialic acid. -
Sequence similarities
Belongs to the immunoglobulin superfamily. SIGLEC (sialic acid binding Ig-like lectin) family.
Contains 4 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 4099 Human
- Entrez Gene: 17136 Mouse
- Entrez Gene: 29409 Rat
- Omim: 159460 Human
- SwissProt: P20916 Human
- SwissProt: P20917 Mouse
- SwissProt: P07722 Rat
- Unigene: 643440 Human
see all -
Alternative names
- GMA antibody
- MAG antibody
- MAG_HUMAN antibody
see all
Images
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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.
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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).
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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.
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All lanes : Anti-MAG/GMA antibody [EPR24276-125] (ab277524) at 1/1000 dilution
Lane 1 : Human cerebellum tissue lysate
Lane 2 : Human liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 69 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?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.
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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.
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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).
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All lanes : Anti-MAG/GMA antibody [EPR24276-125] (ab277524) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse cerebellum tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Rat cerebellum tissue lysate
Lane 5 : Rat liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 69 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?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.
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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).
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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.
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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.
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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).
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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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab277524 has not yet been referenced specifically in any publications.