alpha-Galactosidase A/GLA Antibody [Unconjugated]

Western blot shows lysates of A549 human lung carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human a-Galactosidase A/GLA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6146) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for a-Galactosidase A/GLA at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

a-Galactosidase A/GLA was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human a-Galactosidase A/GLA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6146) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010). LAMP1 was also detected using Mouse Anti-Human LAMP1 Mono-clonal Antibody (Catalog # MAB4800). Cells were stained using the Northern-Lights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009). Cells were counterstained with DAPI (blue). Specific staining was localized to lysosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for alpha ‑Galactosidase A/GLA at approximately 55 kDa (as indicated) using 50 µg/mL of Sheep Anti-Human alpha ‑Galactosidase A/GLA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6146) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

alpha-Galactosidase A/GLA Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human alpha ‑Galactosidase A/GLA
  Leu32-Leu429
  Accession # P06280
• Specificity: Detects human alpha ‑Galactosidase A/GLA in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: GLA
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 50 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for alpha-Galactosidase A/GLA Antibody [Unconjugated]

• agalsidase alfa
• Agalsidase alpha
• Agalsidase
• Alpha-D-galactosidase A
• alpha-D-galactoside galactohydrolase 1
• Alpha-D-galactoside galactohydrolase
• alpha-gal A
• alpha-galactosidase A
• EC 3.2.1
• EC 3.2.1.22
• GALA
• galactosidase, alpha
• GLA
• Melibiase

Background

Human alpha -Galactosidase A is a homodimeric glycoprotein that can release terminal alpha -galactosyl moieties from glycolipids and glycoproteins and catalyze the hydrolysis of melibiose into galactose and glucose (1). It is a lysosomal enzyme and is responsible for degradation of glycolipid globotriaosylceramide (Gb3) (Gal alpha 1‑4Gal beta 1‑4Glc beta ‑ceramide). Mutations in this gene cause Fabry disease, an X-linked hereditary lysosomal storage disease with the accumulation of Gb3 in the walls of small blood vessels, nerves, dorsal root ganglia, renal glomerular and tubular epithelial cells, and cardiomyocytes (2, 3). Inability to prevent the glycosphingolipid deposition can cause hypertension, strokes, heart attack and progressive renal failure (4). Current treatment for Fabry disease is enzyme replacement therapy using intravenously delivered recombinant alpha -Galactosidase A (5, 6).

1. Ioannou, Y.A. et al. (1998) Biochem. J. 332:789.
2. Koide, T. et al. (1990) FEBS Lett. 259:353.
3. Ioannou Y.A, et al. (1992) J. Cell Biol. 119:1137.
4. Germain, D.P. (2002) Expert. Opin. Investig. Drugs. 11:1467.
5. Barngrover, D. (2003) J. Biotechnol. 95:280.
6. Mignani, R. and Cagnoli, L. (2004) J. Nephrol. 17:354.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for alpha-Galactosidase A/GLA Antibody (AF6146)(3)

Publications using AF6146

• Tomsen-Melero, J;Moltó-Abad, M;Merlo-Mas, J;Díaz-Riascos, ZV;Cristóbal-Lecina, E;Soldevila, A;Altendorfer-Kroath, T;Danino, D;Ionita, I;Pedersen, JS;Snelling, L;Clay, H;Carreño, A;Corchero, JL;Pulido, D;Casas, J;Veciana, J;Simó Schwartz, ;Sala, S;Font, A;Birngruber, T;Royo, M;Córdoba, A;Ventosa, N;Abasolo, I;González-Mira, E; Targeted nanoliposomes to improve enzyme replacement therapy of Fabry disease Science advances 2024-12-13 [PMID: 39671483] (ELISA Detection, Rat)
• Malte Lenders, Solvey Pollmann, Melina Terlinden, Eva Brand Pre-existing anti-drug antibodies in Fabry disease show less affinity for pegunigalsidase alfa Molecular Therapy - Methods & Clinical Development 9/8/2022 [PMID: 35990747]
• M Yasuda, MW Huston, S Pagant, L Gan, S St Martin, S Sproul, D Richards, S Ballaron, K Hettini, A Ledeboer, L Falese, L Cao, Y Lu, MC Holmes, K Meyer, RJ Desnick, T Wechsler AAV2/6 Gene Therapy in a Murine Model of Fabry Disease Results in Supraphysiological Enzyme Activity and Effective Substrate Reduction Mol Ther Methods Clin Dev, 2020-07-09;18(0):607-619. 2020-07-09 [PMID: 32775495] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: GLA
• Uniprot:
  • P06280()