Anti-Agrin antibody (ab85174)

Overview

  • Product name
  • Description
    Rabbit polyclonal to Agrin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide from the N terminal residues of Human Agrin protein (NM_198576).

  • Positive control
    • Human skeletal muscle.

Properties

Applications

Our Abpromise guarantee covers the use of ab85174 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/100 - 1/500.

Target

  • Function
    Isoform 1: heparan sulfate basal lamina glycoprotein that plays a central role in the formation and the maintenance of the neuromuscular junction (NMJ) and directs key events in postsynaptic differentiation. Component of the AGRN-LRP4 receptor complex that induces the phosphorylation and activation of MUSK. The activation of MUSK in myotubes induces the formation of NMJ by regulating different processes including the transcription of specific genes and the clustering of AChR in the postsynaptic membrane. Calcium ions are required for maximal AChR clustering. AGRN function in neurons is highly regulated by alternative splicing, glycan binding and proteolytic processing. Modulates calcium ion homeostasis in neurons, specifically by inducing an increase in cytoplasmic calcium ions. Functions differentially in the central nervous system (CNS) by inhibiting the alpha(3)-subtype of Na+/K+-ATPase and evoking depolarization at CNS synapses. This secreted isoform forms a bridge, after release from motor neurons, to basal lamina through binding laminin via the NtA domain.
    Isoform 2: transmembrane form that is the predominate form in neurons of the brain, induces dendritic filopodia and synapse formation in mature hippocampal neurons in large part due to the attached glycosaminoglycan chains and the action of Rho-family GTPases.
    Isoform 1, isoform 4 and isoform 5: neuron-specific (z+) isoforms that contain C-terminal insertions of 8-19 AA are potent activators of AChR clustering. Isoform 5, agrin (z+8), containing the 8-AA insert, forms a receptor complex in myotubules containing the neuronal AGRN, the muscle-specific kinase MUSK and LRP4, a member of the LDL receptor family. The splicing factors, NOVA1 and NOVA2, regulate AGRN splicing and production of the 'z' isoforms.
    Isoform 3 and isoform 6: lack any 'z' insert, are muscle-specific and may be involved in endothelial cell differentiation.
    Agrin N-terminal 110 kDa subunit: is involved in regulation of neurite outgrowth probably due to the presence of the glycosaminoglcan (GAG) side chains of heparan and chondroitin sulfate attached to the Ser/Thr- and Gly/Ser-rich regions. Also involved in modulation of growth factor signaling.
    Agrin C-terminal 22 kDa fragment: this released fragment is important for agrin signaling and to exert a maximal dendritic filopodia-inducing effect. All 'z' splice variants (z+) of this fragment also show an increase in the number of filopodia.
  • Tissue specificity
    Expressed in basement membranes of lung and kidney. Muscle- and neuron-specific isoforms are found. Isoforms (y+) with the 4 AA insert and (z+8) isoforms with the 8 AA insert are all neuron-specific. Isoforms (z+11) are found in both neuronal and non-neuronal tissues.
  • Involvement in disease
    Myasthenic syndrome, congenital, 8
  • Sequence similarities
    Contains 4 EGF-like domains.
    Contains 9 Kazal-like domains.
    Contains 2 laminin EGF-like domains.
    Contains 3 laminin G-like domains.
    Contains 1 NtA (N-terminal agrin) domain.
    Contains 1 SEA domain.
  • Domain
    The NtA domain, absent in TM-agrin, is required for binding laminin and connecting to basal lamina.
    Both laminin G-like 2 (G2) and laminin G-like 3 (G3) domains are required for alpha-dystroglycan/DAG1 binding. G3 domain is required for C-terminal heparin, heparan sulfate and sialic acid binding.
  • Post-translational
    modifications
    Contains heparan and chondroitin sulfate chains and alpha-dystroglycan as well as N-linked and O-linked oligosaccharides. Glycosaminoglycans (GAGs), present in the N-terminal 110 kDa fragment, are required for induction of filopodia in hippocampal neurons. The first cluster (Gly/Ser-rich) for GAG attachment contains heparan sulfate (HS) chains and the second cluster (Ser/Thr-rich), contains chondroitin sulfate (CS) chains. Heparin and heparin sulfate binding in the G3 domain is independent of calcium ions. Binds heparin with a stoichiometry of 2:1. Binds sialic acid with a stoichiometry of 1:1 and binding requires calcium ions.
    At synaptic junctions, cleaved at two conserved sites, alpha and beta, by neurotrypsin. Cleavage at the alpha-site produces the agrin N-terminal 110-kDa subunit and the agrin C-terminal 110-kDa subunit. Further cleavage of agrin C-terminal 110-kDa subunit at the beta site produces the C-terminal fragments, agrin C-terminal 90 kDa fragment and agrin C-terminal 22 kDa fragment. Excessive cleavage at the beta-site releases large amounts of the agrin C-terminal 22 kDa fragment leading to destabilization at the neuromuscular junction (NMJ).
  • Cellular localization
    Cell junction, synapse. Cell membrane and Secreted, extracellular space, extracellular matrix. Synaptic basal lamina at the neuromuscular junction.
  • Information by UniProt
  • Database links
  • Alternative names
    • AGRIN antibody
    • Agrin C-terminal 22 kDa fragment antibody
    • Agrin proteoglycan antibody
    • AGRIN_HUMAN antibody
    • Agrn antibody
    • C22 antibody
    • C90 antibody
    • FLJ45064 antibody
    • OTTHUMP00000044043 antibody
    see all

Images

  • ab85174 staining Agrin in foetal colon tissue by immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) at a dilution of 1/100.

  • ab85174, at 1/100 dilution, staining Agrin in formalin-fixed, paraffin-embedded human skeletal muscle by Immunohistochemistry.

References

ab85174 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat embryonic Dorsal Horn Cells in culture)
Permeabilization
No
Specification
Rat embryonic Dorsal Horn Cells in culture
Fixative
4%PF,0.2% Picric Acid
Username

Ms. Babben Tinner

Verified customer

Submitted Oct 13 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Purified protein (proteoglycan preparation from human milk)
Specification
proteoglycan preparation from human milk
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Serena Leone

Verified customer

Submitted Dec 04 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer & Tris pH 9.0
Sample
Mouse Tissue sections (mouse kidney)
Specification
mouse kidney
Permeabilization
Yes - 0.2 % Triton X in PBS
Fixative
Formaldehyde
Username

Dr. Steffen Rickelt

Verified customer

Submitted Nov 12 2013

The immunogen is a 14 amino acid peptide. It is within this range of amino acids: magrshpgpl rpllpllvva acvlpgaggt cperalerre eeanvvltgt veeilnvdpv.

Thank you for you enquiry regarding ab85174.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase...

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Thank you for contacting Abcam.   I have check with the lab and they have confimred that the immunogen sequence is within the first 1-250aa of the protein. They are unable to provide the actual sequence of the immunogen but I hope that this information...

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