Dystroglycan Antibody [Unconjugated]

Western blot shows lysates of MCF‑7 human breast cancer cell line, SH‑SY5Y human neuroblastoma cell line, human muscle tissue, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for alpha ‑Dystroglycan at approximately 100-160 kDa (as indicated) and beta ‑Dystroglycan at approximately 42-44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Dystroglycan was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to basement membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Dystroglycan knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for alpha ­-Dystroglycan at approximately 110 kDa ­and beta -Dystroglycanat approximately 42 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, IHC, KO
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Dystroglycan Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Dystroglycan
  Gln28-Val749
  Accession # Q14118
• Specificity: Detects human Dystroglycan in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: DAG1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Knockout Validated
  • Western Blot 1 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 Dystroglycan Antibody [Unconjugated]

• A3a
• AGRNR
• DAG1
• Dag-1
• DAG156DAG
• dystroglycan 1 (dystrophin-associated glycoprotein 1)
• Dystroglycan
• Dystrophin-associated glycoprotein 1

Background

Dystroglycan, also DAG-1 (Dystrophin-associated glycoprotein 1) is a 180-200 kDa heterodimeric adhesion molecule that links the cell cytoskeleton to the extracellular matrix. It is found on skeletal muscle, cardiac muscle, fibroblasts, smooth muscle and keratinocytes. DAG-1 binds multiple matrix molecules, including laminin-1 and -2, agrin, and perlecan. Intracellularly, the cytoplasmic tail of DAG-1 contributes to a large 400 kDa complex that interacts with the cytoskeleton. The human DAG-1 preprocursor is a type I transmembrane protein 895 amino acids (aa) in length. It contains a 27 aa signal sequence plus an 868 aa proform that undergoes autocatalysis to generate a 626 aa alpha -chain (aa 28-653), and a 242 aa beta -chain. Mature DAG-1 is a heterodimer composed of noncovalently linked alpha ‑ and beta ‑chains. The alpha -chain possesses one potential Ig-like domain (aa 64-162), a mucin-like region (aa 316-485), and a peptidase S72 domain (aa 500-733). It is O‑glycosylated and runs from 100-160 kDa in SDS-PAGE. The beta -chain is N-glycosylated and runs at 42-44 kDa in SDS-Page. It possesses a short 95 aa extracellular region (aa 654-749) plus a 120 aa cytoplasmic domain (aa 776-895). Membrane cleavage of the beta -chain causes dissociation of the heterodimer and generates a 30 kDa truncated form. Over aa 28-749, human DAG-1 shares 93% aa identity with mouse DAG-1.

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 Dystroglycan Antibody (AF6868)(7)

Publications using AF6868

• P Franzka, SC Schüler, T Kentache, R Storm, A Bock, I Katona, J Weis, K Buder, C Kaether, CA Hübner Impact of Hypermannosylation on the Structure and Functionality of the ER and the Golgi Complex Biomedicines, 2023-01-06;11(1):. 2023-01-06 [PMID: 36672654] (Western Blot, Mouse)
• F Umezawa, M Natsume, S Fukusada, K Nakajima, F Yamasaki, H Kawashima, CW Kuo, KH Khoo, T Shimura, H Yagi, K Kato Cancer Malignancy Is Correlated with Upregulation of PCYT2-Mediated Glycerol Phosphate Modification of alpha-Dystroglycan International Journal of Molecular Sciences, 2022-06-15;23(12):. 2022-06-15 [PMID: 35743105] (Western Blot, Human)
• PT Martin, DA Zygmunt, A Ashbrook, S Hamilton, D Packer, SM Birch, AK Bettis, CJ Balog-Alva, LJ Guo, PP Nghiem, JN Kornegay Short-term treatment of golden retriever muscular dystrophy (GRMD) dogs with rAAVrh74.MHCK7.GALGT2 induces muscle glycosylation and utrophin expression but has no significant effect on muscle strength PLoS ONE, 2021-03-26;16(3):e0248721. 2021-03-26 [PMID: 33770101] (IHC, Canine)
• PJ Lu, JD Tucker, EK Branch, F Guo, AR Blaeser, QL Lu Ribitol enhances matriglycan of &alpha-dystroglycan in breast cancer cells without affecting cell growth Sci Rep, 2020-03-18;10(1):4935. 2020-03-18 [PMID: 32188898] (Western Blot, Human)
• N Gumlaw, LM Sevigny, H Zhao, Z Luo, DS Bangari, E Masterjohn, Y Chen, B McDonald, M Magnay, T Travaline, T Yoshida-Mo, W Fan, D Reczek, JE Stefano, H Qiu, C Beil, C Lange, E Rao, M Lukason, E Barry, WH Brondyk, Y Zhu, SH Cheng biAb Mediated Restoration of the Linkage between Dystroglycan and Laminin-211 as a Therapeutic Approach for alpha-Dystroglycanopathies Mol. Ther., 2019-12-06;0(0):. 2019-12-06 [PMID: 31843448] (Western Blot, Human)
• MP Cataldi, P Lu, A Blaeser, QL Lu Ribitol restores functionally glycosylated ?-dystroglycan and improves muscle function in dystrophic FKRP-mutant mice Nat Commun, 2018-08-27;9(1):3448. 2018-08-27 [PMID: 30150693] (Western Blot, Mouse)
• Structural basis of laminin binding to the LARGE glycans on dystroglycan Nat Chem Biol, 2016-08-15;0(0):. 2016-08-15 [PMID: 27526028] (Western Blot, Rabbit)

Bioinformatics

• Gene Symbol: DAG1
• Uniprot:
  • Q14118(Human)