Antibody Panel useful as specific Marker for the Golgi Apparatus: GM130, GOLGA3, Bet1, 58kD Golgi Protein, TGN38, Mannosidase 2, Membrin, Syntaxin 6 and Colgi Zone

For many research applications it may be useful to be able to stain or to label proteins specific for a certain cell type or cell organell. Antibodies are proven tools for this purpose. They are made to proteins only expressed in the cell type or cell organell of interest.

Acris Antibodies offers a selections of antibodies usefull as organell markers. This “Focus” presents an antibody panel to proteins specific for the Golgi apparatus. These polyclonal and monoclonal antibodies can be used on different species and in most relevant immunological methods, like immunohistochemistry, immunoprecipitation or western blotting. The following protein targets are good candidates for specific staining of the Golgi apparatus by antibodies:

Golgi marker antibody: 58k Golgi protein

58K protein antibodies are excellent for use as markers for the Golgi complex. The 58K protein has been identified as being FTCD, a bifunctional enzyme that channels 1-carbon units from formiminoglutamate, a metabolite of the histidine degradation pathway, to the folate pool.

Golgi marker antibody: GS28/GOS28

GS28, also known as p28 or GOS28, is a 28 kDa integral membrane protein on the surface of the Golgi apparatus that serves as a t- SNARE in ER to Golgi transport. The amino-terminal of GS28/GOS28 is exposed to the cytosol and is anchored to ths cis Golgi via a 20 amino acid carboxyl-terminal hydrophobic tail. GS28 co-immunoprecipitates complexes consisting of syntaxin 5, rbet1, membrin, rsec22, and rsly1, and is therefore implicated in ERto- Golgi or intra-Golgi vesicle transport. ATP hydrolysis by NSF in the presence of SNAP causes the dissociation of GOS28 and syntaxin 5 complex.

Golgi marker antibody: TGN38

Recognizes a glycoprotein designated TGN38 that is found primarily in the Trans-Golgi network and has been found to be an excellent marker for this cellular organelle. TGN38 is likely to have a role in intracellular transport

Golgi marker antibody: Mannosidase 2

Mannosidase 2 is a 135kD protein located on the luminal side of the Golgi membranes. It is useful as an immunocytochemical marker for the Golgi apparatus as well as for studying the fate of the Golgi complex in dividing cells.

Golgi marker antibody: Bet1

Bet1 is a 17 kDa transmembrane protein that mediates protein transport between endoplasmic reticulum (ER) and the Golgi apparatus. rBet1 serves as the vesicle SNAP receptor (v-SNARE) on ER-derived vesicles, which interacts with its partner t-SNARE on the Golgi apparatus during vesicle docking and fusion. rBet1 is found in a complex with syntaxin 5, GS27/membrin, GOS28, rsec22 and rsly1. Antibodies to rBet1 have been shown to inhibit the transfer of vesicular stomatitis virus G-protein from the ER to the Golgi apparatus in a cell free assay, providing evidence for the involvement of rBet1 in ER to Golgi transport.

Golgi marker antibody: Syntaxin 6

Syntaxin 6 is a 35 kDa type II integral membrane protein with two coiled-coil domains and a carboxyl-terminal membrane anchor. Immunoelectron microscopy studies show that syntaxin 6 is primarily localized to the trans-Golgi network (TGN), and is partially co-localized with AP-1 on clathrincoated membranes. During secretory granule maturation, syntaxin 6 and MPR-ligand complexes are postulated to be transferred from immature granules to endosomes by clathrincoated vesicles, as evidenced by immunoelectron microscopy . These results suggest that syntaxin 6 mediates a TGN trafficking targeting endosome.

Golgi marker antibody: GM130/GOLGA2

The Golgi apparatus, which participates in glycosylation and transport of proteins and lipids in the secretory pathway, consists of a series of stacked cisternae (flattened membrane sacs). Interactions between the Golgi and microtubules are thought to be important for the reorganization of the Golgi after it fragments during mitosis. GM130 encodes a member of the Golgin protein family and is localised to the Golgi. The protein encoded by GM130 has been postulated to play roles in the stacking of Golgi cisternae, maintaining cis-Golgi structure and in vesicular transport. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of these variants has not been determined. GM130 is part of a larger oligomeric complex which interacts with p115, GORASP1/GRASP65 and RAB1B that has been activated by GTP-binding. GM130 is useful as a cis-Golgi marker in immunohistochemistry.

Golgi marker antibody: Golgin-160/GOLGA3

By screening a human liver carcinoma cDNA expression library with serum from a systemic lupus erythematosus patient, cDNAs were cloned representing 2 genes, GOLGA2 and GOLGA3, that encode Golgi complex antigens. The GOLGA3 cDNAs isolates contains a partial coding sequence. The protein product of the GOLGA3 gene was designated golgin-160 based on its 160-kD molecular mass in mammalian cell extracts. Antibodies specific to golgin-160 shows anti-Golgi reactivity by immunofluorescence; the Golgi staining is blocked by treatment with brefeldin A (BFA), a fungal antibiotic known to disrupt the stacked structure of the Golgi complex and to affect Golgi coatomer proteins.

Golgi marker antibody: Membrin/GS27

Membrin, also known as GS27 (Golgi SNARE of 27K), is a 27kD integral membrane protein on the surface of the Golgi apparatus that serves as a t-SNARE in ER to Golgi transport. GS27 is a member of a protein complex that consists of cis-Golgi vesicle receptor syntaxin 5 and GS28, v-SNARE rbet1 and rsec22, and rsly1. Immunoprecipitation analysis has shown that there are two subcomplexes within this complex: one contains syntaxin 5 and GOS28 and the other one contains syntaxin 5, GS27/membrin, rsec22 and rbet1. These complexes are postulated to direct the fusion of ER-derived vesicles with Vesicular tubular cluster (VTC), and the fusion of VTCs to form cis-Golgi compartment.

Staining pictures:

Fig.1: Golgi marker antibody to GOLGA3 cat# EB06926 (0.1µg/ml) staining of Hela lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence

Fig.2: Golgi marker antibody to 58k Golgi protein cat#  EB06442 (0.03µg/ml) staining of Human Liver lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence

Fig.3: Immunofluorescence staining of TGN-38 in NRK cells using Golgi marker antibody cat# NB300-575

Golgi markers antibody

58kD Golgi protein antibody, GS28 antibody, TGN38 antibody, Mannosidase 2 antibody, Bet1 antibody, Syntaxin 6 antibody, GM130/GOLGA2 antibody, Golgin-160/GOLGA3 antibody and Colgi Zone antibody

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