Anti-58K Golgi protein antibody [58K-9] - Golgi Marker

5 product images

• 

  Immunocytochemistry - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043)

  ab27043 staining 58K Golgi protein - Golgi Marker in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), 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 ab27043 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Image courtesy of Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

  Immunocytochemistry - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043)

  ab27043 (1µg/ml, 5µg/ml and 10µg/ml) staining 58K Golgi protein in SK-N-SH cells (green). Cells were fixed in Methanol, permabilised using 0.5% Triton X100 in PBS and counterstained with DAPI in order to highlight the nucleus (red).

• 

  Western blot - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043)

  Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  All lanes: Western blot - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043) at 1 µg/mL

  Lane 1: Liver (Rat) Tissue Lysate, blocked with 5% BSA at 20 µg

  Lane 2: Liver (Rat) Tissue Lysate, blocked with 3% Milk at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 58 kDa

  Exposure time: 3min

• 

  Western blot - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043)

  Western blot: Anti-FTCD antibody [58K-9] (ab27043) staining at 2.5 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab27043 was shown to bind specifically to FTCD. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043) at 2.5 µg/mL

  Lane 1: Human Liver cell lysate at 30 µg

  Lane 2: HepG2 cell lysate at 30 µg

  Lane 3: Rat Liver cell lysate at 30 µg

  Lane 4: Human Brain cell lysate at 30 µg

  Lane 5: SH-SY5Y cell lysate at 30 µg

  Lane 6: HeLa cell lysate at 30 µg

  Secondary

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

  Performed under reducing conditions.

  Observed band size: 58 kDa

• 

  Flow Cytometry (Intracellular) - Anti-58K Golgi protein antibody [58K-9] - Golgi Marker (ab27043)

  Overlay histogram showing HepG2 cells stained with ab27043 (red line). The cells were PFA-fixed and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab27043, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.