Anti-BAP31 antibody [7A3BB6]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-BAP31 antibody [7A3BB6] (AB112993)

ab112993 at 5 μg/ml staining BAP31 in Human fibroblast cells by Immunocytochemistry (4% paraformaldehyde fixed and 0.1% Triton X-100 permeabilized) followed by Alexa Fluor^® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour (green). The target locates mainly in the endoplasmic reticulum.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BAP31 antibody [7A3BB6] (AB112993)

IHC image of BAP31 staining in Human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab112993, 5μg/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.

• Flow Cyt

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Flow Cytometry - Anti-BAP31 antibody [7A3BB6] (AB112993)

ab112993 at 1ug/ml staining BAP31 in MeOH-fixed HL60 cells by Flow Cytometry (blue). Isotype control antibody (red).

• WB

Lab

Western blot - Anti-BAP31 antibody [7A3BB6] (AB112993)

Lanes 1-4 : Merged signal (red and green). Green - ab112993 observed at 28 kDa. Red - loading control ab52901 observed at kDa.

ab112993 Anti-BAP31 antibody [7A3BB6] was shown to specifically react with BAP31 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266634 (knockout cell lysate ab257857) was used. Wild-type and BAP31 knockout samples were subjected to SDS-PAGE. ab112993 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BAP31 antibody [7A3BB6] (ab112993) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

BCAP31 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human BCAP31 (BAP31) knockout HEK-293T cell line (<a href='/ko/products/cell-lines/human-bcap31-bap31-knockout-hek-293t-cell-line-ab266634'>ab266634</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

NCCIT cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

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• WB

Unknown

Western blot - Anti-BAP31 antibody [7A3BB6] (AB112993)

All lanes:

Western blot - Anti-BAP31 antibody [7A3BB6] (ab112993) at 0.5 µg/mL

Lane 1:

HepG2 Cell Lysate

Lane 2:

H9C2 Cell Lysate

Lane 3:

MEF Cell Lysate

Predicted band size: 28 kDa

false