I am searching for a dystroclycan antibodiy workin in IF and WB analysis.We had one, but it did not work out in my hands (I tried really a lot). Now I am looking for a new one (commercial).
Hi Isabell,
I'm working with dystroglycan in my PhD. Do you want an antibody for glycosylation of alpha-dystroglycan or for the core of the protein?.
For glycosylation, IIH6 (from Millipore) is the best that i have tried.
For core of alpha, i'm not lucky. I have tried several of them (commercial and one non-commercial) and i haven't obtained results.
Hi,
I have no preference. Since I am interested to show the dystroglycan distribution on Schwann cells (there might be differences in my knockout mice), The antibody should work in the peripheral nervous system isoforms of dystroglycan. I just read that there is an antibody available for the beta subunit. Do yo have any experience with that one?
Hi again,
If only you want to see distribution, beta-dystroglycan may be the best choice. We use anti-beta dystroglycan from Novocastra but i see that the same antibody is distributed by others (the name i think is "clone 43DAG1/8D5").
I've used this antibody in immunohistofluorescence, mainly in muscle tissue and it works very well. In nervous system, I don't know how it works, sorry; but is a good antibody, so the differences should be minimal.
Hello,
You may check a free antibody database available at labome.com. The information for dystroglycan antibodies is provided here: https://www.labome.com/review/gene/human/DAG1-antibody.html. Santa Cruz Biotechnology has mouse monoclonal (7D11) sc-33701 ab suitable for WB (ref: Gerin et al., Nat Commun. 2016), rabbit polyclonal (H-300) sc-28534 ab suitable for WB and IHC (ref: Noel et al., PLoS One. 2015), and goat polyclonal (C-20) sc-16165 ab (Ogasawara et al., Physiol Rep. 2014). Also, EMD Millipore offers mouse monoclonal (IIH6C4) DAG1 antibody (Merck Millipore, 05-593) (ref: Tjondrokoesoemo et al., Hum Mol Genet. 2016) and (VIA4-1) 05-298 (Domi et al., Skelet Muscle. 2015) abs suitable for both WB and IHC.
Hello,
Did you solve your problem? I encountered the same problem now. I try to see the expression level of alpha-dystroglycan in breast cancer cell lines BT-549 and MDA-MB-231, but unfortunately, I got no clear bands in WB.
I have tried IIH6 and VIA41 (Santa-Cruze) ，still no positive results.
Hello to everybody,
I have tested IIH6 ab with C2C12 cells and fibroblasts.
You need to use excess amount of ab and load more than 70 ug of protein per well. Since alpha dystroglycan is an extracellular protein cells should be collected by scraping (not with trypsinization).
The source of the IIH6 was Developmental Studies Hybridoma Bank.
http://dshb.biology.uiowa.edu/dystroglycan?sc=18&category=9199
They have also VIA41 antibody but it did not work for me.
SCBT has an antibody for core alpha dyst.
https://www.scbt.com/scbt/product/alpha-dystroglycan-antibody-d-3
It gave me a good strong sharp band. However, the identity of the band needs to be verified. I have transfected DAG1 gene to my cells but did not see any up regulation (confusing). So I need to verify it with a knockdown experiment.
EDIT: I found that SCBT D3 ab is very good at detecting 37 kDa secreted form of alpha DG. It gives very strong band. Indeed, this ab was raised against N terminal end of the protein. Note that it is secreted, not in cell lysates. You dont get any band at 37 kDa with cell lysates.
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