Anti-ARF1 antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 antibody (AB183576)

Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue staining ARF1 with ab183576 at 1/100 dilution (right), and negative control (left) with no primary antibody. Atigen retrieval method using sodium citrate pH6, and detection was with an HRP-conjugated secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ARF1 antibody (AB183576)

Immunofluorescent analysis of HeLa cells (formalin-fixed, 0.1% Triton X-100 permeabilized) labeling ARF1 with ab183576 at 1/100 dilution followed with DyLight 488 goat anti-rabbit IgG secondary antibody at 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 antibody (AB183576)

Immunohistochemistry analysis of paraffin-embedded human colon carcinoma tissue staining ARF1 with ab183576 at 1/100 dilution (right), and negative control (left) with no primary antibody. Atigen retrieval method using sodium citrate pH6, and detection was with an HRP-conjugated secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ARF1 antibody (AB183576)

Immunocytochemistry analysis of 4% paraformaldehyde-fixed 0.1% Triton™ X-100 permeabilized MDA-MB-231 cells staining ARF1 with ab183576 at 5μg/ml, and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution (green). Nuclear counterstain was ProLong™ Diamond Antifade Mountant with DAPI (blue), and F-actin was stained with Rhodamine Phalloidin (red). Negative control used no primary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARF1 antibody (AB183576)

Immunohistochemistry analysis of paraffin-embedded human mouse colon tissue staining ARF1 with ab183576 at 1/100 dilution (right), and negative control (left) with no primary antibody. Atigen retrieval method using sodium citrate pH6, and detection was with an HRP-conjugated secondary antibody.

• IP

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Immunoprecipitation - Anti-ARF1 antibody (AB183576)

Detection of ARF1 in Immunoprecipitates of HeLa cell lysate. Antigen-antibody complexes were formed by incubating 500μg of HeLa whole cell lysate with 2μg ab183576 (lane 2) compared with HeLa cell lysate as a positive control (lane 1). For detection, ab183576 was used at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-ARF1 antibody (ab183576)

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-ARF1 antibody (AB183576)

Lanes 1 - 4 : Merged signal (red and green). Green - ab183576 observed at 18 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab183576 was shown to react with ARF1 in wild-type HeLa cells in Western blot with loss of signal observed in ARF1 knockout cell line ab264939 (ARF1 knockout cell lysate ab257353). Wild-type HeLa and ARF1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab183576 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ^°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ARF1 antibody (ab183576) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ARF1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ARF1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-arf1-knockout-hela-cell-line-ab264939'>ab264939</a>)

Lane 3:

MDA-MB-231 cell lysate at 20 µg

Lane 4:

PANC-1 cell lysate at 20 µg

Predicted band size: 21 kDa

Observed band size: 18 kDa

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• WB

Supplier Data

Western blot - Anti-ARF1 antibody (AB183576)

All lanes:

Western blot - Anti-ARF1 antibody (ab183576) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 25 µg

Lane 2:

U2 OS cell lysate at 25 µg

Lane 3:

3T3 cell lysate at 25 µg

Lane 4:

NRK cell lysate at 25 µg

Lane 5:

MDCK cell lysate at 25 µg

Secondary

All lanes:

goat anti-rabbit IgG-HRP at 1/1500 dilution

Predicted band size: 21 kDa

true

• WB

Supplier Data

Western blot - Anti-ARF1 antibody (AB183576)

All lanes:

Western blot - Anti-ARF1 antibody (ab183576) at 1/3000 dilution

Lane 1:

MDA-MB-231 whole cell extract (human breast adenocarcinoma cell line) at 30 µg

Lane 2:

MDA-MB-231 whole cell extract (human breast adenocarcinoma cell line) treated with 5ug/ml Brefeldin for 60 minutes at 30 µg

Lane 3:

HeLa whole cell extract at 30 µg

Lane 4:

U2 OS whole cell extract (human bone osteosarcoma epithelial cell line) at 30 µg

Lane 5:

DU 145 whole cell extract (Human prostate carcinoma cell line) at 30 µg

Lane 6:

PC-3 whole cell extract (human prostate cancer cell line) at 30 µg

Lane 7:

SK-OV-3 whole cell extract (Human ovarian cancer cell line) at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/4000 dilution

Predicted band size: 21 kDa

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