Anti-MAG Picoband™ Antibody (monoclonal, 2G11)

Product Name

Anti-MAG Picoband™ Antibody (monoclonal, 2G11)

View all MAG/Siglec-4a Antibodies

SKU/Catalog Number

M03019

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-MAG Picoband™ Antibody (monoclonal, 2G11) catalog # M03019. Tested in Flow Cytometry, IF, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-MAG Picoband™ Antibody (monoclonal, 2G11) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M03019)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Monoclonal

Clone Number

2G11

Isotype

Mouse IgG2a

Immunogen

E.coli-derived human MAG recombinant protein (Position: E34-R605).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M03019 is reactive to MAG in Human, Mouse, Rat

Applications

M03019 is guaranteed for Flow Cytometry, IF, IHC, WB Boster Guarantee

Observed Molecular Weight

100 kDa

Calculated molecular weight

69.069kDa

Background of MAG/Siglec-4a

MAG (Myelin-associated glycoprotein),also known as SIGLEC4A, is a cell membrane glycoprotein that is a member of the SIGLEC family of proteins and is a functional ligand of the NOGO-66 receptor, NgR. It is though to be involved in the process of myelination. MAG is a sialic acid-binding SIGLEC protein and is a functional ligand for the NOGO receptor.The MAG gene is mapped on 19q13.12. Cleavage of GPI-linked proteins from axons protects growth cones from MAG-induced collapse, and dominant-negative NgR eliminates MAG inhibition of neurite outgrowth. MAG-resistant embryonic neurons were rendered MAG-sensitive by expression of NgR. MAG binds specifically to an NgR-expressing cell line in a GPI-dependent and sialic acid-independent manner. Experiments blocking NgR from interacting with MAG prevented inhibition of neurite outgrowth by MAG. In cultured human embryonic kidney (HEK) cells expressing the NOGO receptor, p75 (NTR) was required for MAG-induced intracellular calcium elevation.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse,Rat
Immunofluorescence, 5 μg/ml, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of MAG using anti-MAG antibody (M03019).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-MAG antigen affinity purified monoclonal antibody (Catalog # M03019) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for MAG at approximately 100KD. The expected band size for MAG is at 100KD.

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Figure 2. IHC analysis of MAG using anti-MAG antibody (M03019).
MAG was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-MAG Antibody (M03019) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 3. IHC analysis of MAG using anti-MAG antibody (M03019).
MAG was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-MAG Antibody (M03019) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of U87 cells using anti-MAG antibody (M03019).
Overlay histogram showing U87 cells stained with M03019 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-MAG Antibody (M03019, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 5. IF analysis of GFAP and MAG using anti-GFAP antibody and anti-MAG antibody (M03019).
GFAP and MAG was detected in a paraffin-embedded section of rat cerebellum tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-GFAP antibody and mouse anti-MAG antibody (M03019) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142), DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For MAG (Source: Uniprot.org, NCBI)

Alternative Names

GMAsialic acid binding Ig-like lectin 4A; MAG; myelin associated glycoprotein; myelin-associated glycoprotein; sialic acid-binding immunoglobulin-like lectin 4A; Siglec4a; Siglec-4a; S-MAG MAG GMA, S-MAG, SIGLEC-4A, SIGLEC4A, SPG75 myelin associated glycoprotein myelin-associated glycoprotein|sialic acid binding Ig-like lectin 4A|sialic acid-binding immunoglobulin-like lectin 4A

For more info on MAG, check out the MAG Infographic

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In this infographic, you will see the following information for MAG: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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