Confocal immunofluorescent analysis of A-172 cells (left) and HeLa cells (right) using GM130 (D6B1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate). Actin filaments were labeled with DyLight™ 488 Phalloidin #12935 (green) prior to mounting in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated GM130 (D6B1) XP® Rabbit mAb #12480.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
GM130 (D6B1) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total GM130 protein. This antibody may cross-react with a protein of unknown origin at 30 kDa.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr185 of human GM130 protein.
The Golgi apparatus functions in the modification, organization, and transport of proteins and membranes targeted to other parts of the cell, such as the plasma membrane, lysosomes, and endosomes. This regulated transport is important for appropriate protein localization, secretion, and signal transduction. Members of the Golgin family of proteins, including GM130, Giantin, p115, and GRASP65, are defined by their presence in the Golgi matrix and by their long coiled coil domains. Golgin function, which is regulated in part by small GTPases of the Rab and Arl families, includes establishing and maintaining Golgi structure and transport (reviewed in 1). The Golgi cisternae are stacked and linked laterally to form a ribbon. GRASP65 and GM130 are required for membrane fusion events that mediate ribbon formation during Golgi assembly. These lateral fusion events allow for uniform distribution of Golgi enzymes (2). GM130 and Giantin interact with the transport factor p115 to facilitate endoplasmic reticulum (ER)-Golgi transport (3). GM130 is also involved in the transport of the Ether-a-go-go-Related (hERG) potassium ion channel. Inappropriate hERG localization may be an underlying cause in Long QT syndrome, a hereditary and potentially fatal cardiac arrhythmia (4). Further, GM130 was implicated in signal transduction regulating invasion, migration, and cell polarization via its interaction with and activation of serine/threonine kinases YSK1 and Mst4 (5).
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