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Alpha-dystroglycan Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER1901-72

Alpha-dystroglycan Rabbit Polyclonal Antibody

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Alpha-dystroglycan Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within Human Alpha-dystroglycan aa 15-64 / 895.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 97 kDa

Positive Control

Neuro-2a cell lysate, C6 cell lysate, mouse heart tissue lysate, rat heart tissue lysate, SkBr3, Rat skeletal muscle tissue, human placenta tissue, Mouse heart tissue, A431.

Conjugation

unconjugated

RRID

AB_3069362

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB
human
Not Recommended
mouse
Tested Tested
rat
Tested Tested

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:50-1:200

  • IF-Cell

  • 1:50-1:200

  • FC

  • 1:50-1:200

Target

Function

Dystroglycan (DG) is a cell surface receptor for several extracellular matrix molecules including laminins, Agrin and Perlecan. Dystroglycan function is required for the formation of basement membranes in early development and the organization of Laminin on the cell surface. α-dystroglycan is a membrane- associated, extracellular glycoprotein that is anchored to the cell-membrane by binding to the transmembrane glycoprotein α-dystroglycan to form an α/β-dystroglycan-complex. Additionally, dystroglycan is part of a multimolecular complex, where it associates with dystrophin, at the sarcolemma, to form the dystrophin-associated protein complex, or with utrophin, at the neuromuscular junction, to form the utrophin-associated protein complex. Dystroglycan is also thought to participate in the clustering of nicotinic acetylcholine receptors at the neuromuscular junction.

Background References

1. Traverso M et al. DAG1 haploinsufficiency is associated with sporadic and familial isolated or pauci-symptomatic hyperCKemia. Eur J Hum Genet. 2024 Mar

2. Yao J et al. Fat Mass- and Obesity-Associated Protein (FTO) Promotes the Proliferation of Goat Skeletal Muscle Satellite Cells by Stabilizing DAG1 mRNA in an IGF2BP1-Related m(6)A Manner. Int J Mol Sci. 2024 Sep

Tissue Specificity

Expressed in a variety of fetal and adult tissues. In epidermal tissue, located to the basement membrane. Also expressed in keratinocytes and fibroblasts.

Post-translational Modification

O-glycosylated. POMGNT1 catalyzes the initial addition of N-acetylglucosamine, giving rise to the GlcNAc(beta1-2)Man(alpha1-)O-Ser/Thr moiety and thus providing the necessary basis for the addition of further carbohydrate moieties. Alpha-dystroglycan is heavily O-glycosylated comprising of up to two thirds of its mass and the carbohydrate composition differs depending on tissue type. Mucin-type O-glycosylation is important for ligand binding activity. O-mannosylation of alpha-DAG1 is found in high abundance in both brain and muscle where the most abundant glycan is Sia-alpha-2-3-Gal-beta-1-4-Glc-NAc-beta-1-2-Man. In muscle, glycosylation on Thr-317, Thr-319 and Thr-379 by a phosphorylated O-mannosyl glycan with the structure 2-(N-acetylamido)-2-deoxygalactosyl-beta-1,3-2-(N-acetylamido)-2-deoxyglucosyl-beta-1,4-6-phosphomannose is mediated by like-acetylglucosaminyltransferase (LARGE1) protein and is required for laminin binding. O-mannosylation is also required for binding lymphocytic choriomeningitis virus, Old World Lassa fever virus, and clade C New World arenaviruses. The O-glycosyl hexose on Thr-367, Thr-369, Thr-372, Thr-381 and Thr-388 is probably mannose. O-glycosylated in the N-terminal region with a core 1 or possibly core 8 glycan.; The beta subunit is N-glycosylated.; Autolytic cleavage produces the alpha and beta subunits. In cutaneous cells, as well as in certain pathological conditions, shedding of beta-dystroglcan can occur releasing a peptide of about 30 kDa.; SRC-mediated phosphorylation of the PPXY motif of the beta subunit recruits SH2 domain-containing proteins, but inhibits binding to WWW domain-containing proteins, DMD and UTRN. This phosphorylation also inhibits nuclear entry.

Subcellular Location

Secreted, extracellular space; Cell membrane, Cytoplasm, cytoskeleton, Nucleus, nucleoplasm, sarcolemma, Postsynaptic cell membrane.

UNIPROT

SwissProt: Q14118 Human

SwissProt: Q62165 Mouse

Entrez Gene: 114489 Rat

Synonyms

156DAG antibody

A3a antibody

Agrin receptor antibody

AGRNR antibody

Alpha-DG antibody

alpha-DG-N antibody

Beta-DG antibody

Beta-dystroglycan antibody

DAG antibody

Dag1 antibody

Expand

Images

  • Western blot analysis of Alpha-dystroglycan on different lysates with Rabbit anti-Alpha-dystroglycan antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>) at 1/5,000 dilution.<br /><br />Lane 1: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 2: C6 cell lysate (20 µg/Lane)<br />Lane 3: Mouse heart tissue lysate (40 µg/Lane)<br />Lane 4: Rat heart tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 97 kDa<br />Observed band size: 97 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Alpha-dystroglycan on different lysates with Rabbit anti-Alpha-dystroglycan antibody (ER1901-72) at 1/5,000 dilution.

    Lane 1: Neuro-2a cell lysate (20 µg/Lane)
    Lane 2: C6 cell lysate (20 µg/Lane)
    Lane 3: Mouse heart tissue lysate (40 µg/Lane)
    Lane 4: Rat heart tissue lysate (40 µg/Lane)

    Predicted band size: 97 kDa
    Observed band size: 97 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1901-72) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Alpha-dystroglycan in SkBr3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Alpha-dystroglycan in SkBr3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-72, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded Mouse heart tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded Mouse heart tissue using anti-Alpha-dystroglycan antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Alpha-dystroglycan was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ER1901-72" style="font-weight: bold;text-decoration: underline;">ER1901-72</a>, 1/100) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

    Flow cytometric analysis of Alpha-dystroglycan was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-72, 1/100) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

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