Anti-Golgin 97抗体(ab84340)

ab84340 staining Golgin 97 in HeLa cells. The cells were fixed with 100% Methanol (5min), permeabilized with 0.1%PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab84340 at 5μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

 

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

IHC image of Golgin 97 antibody staining in a section of formalin-fixed paraffin-embedded normal human pancreas* performed on a Leica BOND^TM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab84340, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

hCMEC/D3 cells stained for Golgin 97 (green) using ab84340 at 1/100 dilution in ICC/IF, followed by Alexa Fluor 488^® congugated Goat Anti-Rabbit IgG.

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ICC/IF image of ab84340 stained human HepG2 cells. The cells were methanol fixed (5 min), permeabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab84340, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

ab84340 staining Golgin 97 in murine testis tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 0.01M sodium citrate. Samples were then blocked using 10%FCS, 0.5% BSA, 1% Triton X-100, 88.5% PBS X1 and then incubated with ab84340 at a 1/100 dilution for 10 hours at 4°C. The secondary used was an Alexa-Fluor 594 conjugated goat anti-rabbit IgG used at a 1/200 dilution.

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ab84340 staining Golgin-97 in MCF7 cells treated with brefeldin A (ab120299), by ICC/IF. Increase in Golgin-97 expression correlates with increased concentration of brefeldin A, as described in literature.
The cells were incubated at 37øC for 1.5 h in media containing different concentrations of ab120299 (brefeldin A) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab84340 (5 µg/ml) was performed overnight at 4øC in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.