Anti-Golgin 97 antibody [EPR27333-18]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Golgin 97 with ab308520 at 1/250 (1.828 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing the staining co-located with Golgi marker in HeLa cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Golgin 97 with ab308520 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Golgin 97 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab308520 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308520 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab308520 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308520 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 24 seconds

All lanes:

Immunoprecipitation - Anti-Golgin 97 antibody [EPR27333-18] (ab308520) at 1/1000 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 97 kDa

false

Exposure time: 24s

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (mouse myoblast) cells labelling Golgin 97 with ab308520 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labelling Golgin 97 with ab308520 at 1/250 (1.828 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing the staining co-located with Golgi marker in C2C12 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• WB

Supplier Data

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (ab308520) at 1/1000 dilution

Lane 1:

Hela (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate, at 20 µg

Lane 2:

Hela transfected with siRNA specifically targeti Golgin 97 whole cell lysate, at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 97 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Blocking and diluting buffer and concentration : 5% NFDM/TBST  Exposure time : Lane 1 and 2 : 37seconds, Lane 3-5 : 180 seconds.

All lanes:

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (ab308520) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

C2C12 (mouse myoblast) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell)whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 97 kDa

false

• WB

Lab

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Western blot : Anti-GOLGA1 antibody [EPR27333-18] (ab308520) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab308520 was shown to bind specifically to GOLGA1. A band was observed at 99 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in GOLGA1 knockout cell line. To generate this image, wild-type and GOLGA1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (ab308520) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human GOLGA1 Knockout HCT116 cell line (<a href='/ko/products/cell-lines/human-golga1-knockout-hct116-cell-line-ab324223'>ab324223</a>) at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Ramos cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 99 kDa

false

• WB

Supplier Data

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (AB308520)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 92 seconds

All lanes:

Western blot - Anti-Golgin 97 antibody [EPR27333-18] (ab308520) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 88 kDa

Observed band size: 97 kDa

false

Exposure time: 92s