Anti-MGEA5/OGA Antibody Picoband™

Product Name

Anti-MGEA5/OGA Antibody Picoband™

View all O-GlcNAcase/OGA/MGEA5 Antibodies

SKU/Catalog Number

A32463

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-MGEA5/OGA Antibody Picoband™ catalog # A32463. Tested in Flow Cytometry, 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-MGEA5/OGA Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A32463)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at C-terminus of human OGA, identical to the related mouse and rat sequences.

*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

A32463 is reactive to OGA in Human, Mouse, Rat

Applications

A32463 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

130 kDa

Calculated molecular weight

102.915kDa

Background of O-GlcNAcase/OGA/MGEA5

Protein O-GlcNAcase (EC 3.2.1.169, OGA, glycoside hydrolase O-GlcNAcase, O-GlcNAcase, BtGH84, O-GlcNAc hydrolase) is an enzyme with systematic name (protein)-3-O- (N-acetyl-D-glucosaminyl)-L-serine/threonine N-acetylglucosaminyl hydrolase. The dynamic modification of cytoplasmic and nuclear proteins by O-linked N-acetylglucosamine (O-GlcNAc) addition and removal on serine and threonine residues is catalyzed by OGT (MIM 300255), which adds O-GlcNAc, and MGEA5, a glycosidase that removes O-GlcNAc modifications.

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, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of MGEA5/OGA using anti-MGEA5/OGA antibody (A32463).
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 30 ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: mouse brain tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MGEA5/OGA antigen affinity purified polyclonal antibody (Catalog # A32463) at 0.5 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MGEA5/OGA at approximately 130 kDa. The expected band size for MGEA5/OGA is at 103 kDa.

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Figure 2. IHC analysis of MGEA5/OGA using anti-MGEA5/OGA antibody (A32463).
MGEA5/OGA was detected in a paraffin-embedded section of rat spleen 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 2 μg/ml rabbit anti-MGEA5/OGA Antibody (A32463) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

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Figure 3. IF analysis of MGEA5/OGA using anti-MGEA5/OGA antibody (A32463).
MGEA5/OGA was detected in an immunocytochemical section of T-47D cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-MGEA5/OGA Antibody (A32463) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.

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Figure 4. Flow Cytometry analysis of U87 cells using anti-MGEA5/OGA antibody (A32463).
Overlay histogram showing U87 cells stained with A32463 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MGEA5/OGA Antibody (A32463, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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

Alternative Names

bifunctional protein NCOAT; FLJ11229; GH84; HEXC; HEXC3; Hexosaminidase B; hyaluronidase in meningioma; KIAA0679; MEA5FLJ23355; meningioma expressed antigen 5 (hyaluronidase); Meningioma-expressed antigen 5; MGEA5; NCOAT; Nuclear cytoplasmic O-GlcNAcase and acetyltransferase; OGA; OGlcNAcase; O-GlcNAcase OGA MEA5, MGEA5, NCOAT O-GlcNAcase protein O-GlcNAcase|N-acetyl-beta-D-glucosaminidase|N-acetyl-beta-glucosaminidase|beta-N-acetylglucosaminidase|beta-N-acetylhexosaminidase|beta-hexosaminidase|bifunctional protein NCOAT|hyaluronidase in meningioma|meningioma expressed antigen 5 (hyaluronidase)|meningioma-expressed antigen 5|nuclear cytoplasmic O-GlcNAcase and acetyltransferase

For more info on OGA, check out the OGA Infographic

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In this infographic, you will see the following information for OGA: 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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