Anti-ARF1 antibody

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-ARF1 antibody (AB58578)

Flow cytometric analysis of paraformaldehyde fixed A431 cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10µg/mL) followed by Alexa Fluor 488 secondary antibody (1µg/mL). IgG control : Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

• WB

Lab

Western blot - Anti-ARF1 antibody (AB58578)

Lanes 1 - 2 : Merged signal (red and green). Green - ab58578 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab58578 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 ab58578 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Donkey anti-Goat IgG H&L (IRDye^® 800CW) preabsorbed (ab216775) and Donkey anti-Mouse 680RD secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ARF1 antibody (ab58578) at 1 µg/mL

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>)

Predicted band size: 21 kDa

Observed band size: 18 kDa

false

• WB

Supplier Data

Western blot - Anti-ARF1 antibody (AB58578)

Lysates in RIPA buffer. Detected by chemiluminescence.

Lane 1:

Western blot - Anti-ARF1 antibody (ab58578) at 1 µg/mL

Lanes 2 - 3:

Western blot - Anti-ARF1 antibody (ab58578) at 0.03 µg/mL

Lane 1:

A431 cell lysate at 35 µg

Lane 2:

HeLa cell lysate at 35 µg

Lane 3:

MCF7 cell lysate at 35 µg

false

• WB

Supplier Data

Western blot - Anti-ARF1 antibody (AB58578)

Lysates in RIPA buffer. Detected by chemiluminescence.

All lanes:

Western blot - Anti-ARF1 antibody (ab58578) at 1 µg/mL

Lane 1:

NIH3T3 cell lysate at 35 µg

Lane 2:

KNRK cell lysate at 35 µg

Lane 3:

MDCK cell lysate at 35 µg

false

• WB

CiteAb

Western blot - Anti-ARF1 antibody (AB58578)

ARF1 western blot using anti-ARF1 antibody ab58578. Publication image and figure legend from Singh, V., Davidson, A. C., et al., 2020, Int J Mol Sci, PubMed 32252226.

ab58578 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab58578 please see the product overview.

(A) Microscopy images depicting phagocytosis of labelled E. coli particles by PMA differentiated THP-1 macrophages (control) or upon silencing ArcC4, Arf1, Arf3, Arf1& Arf3, or Arf6 using siRNA. Internalized bacteria are shown in red while actin is stained using phalloidin (green). (B) Quantification of the phagocytosed E. coli by THP-1 macrophages as described in (A). Phagocytosis of phRodo-conjugated E. coli particles in THP-1 macrophages (control) or upon using actin inhibitor (CytoD), Arno inhibitor (SecinH3), or silencing of Arf1 or Arf6 in Arno-inhibited cells. Scale bar 10 μm (D). Represents quantification of the percentage of internalized bacteria under conditions as described for (C). (E) Immunoblot confirming the silencing of the mentioned proteins using SiRNA. Each bar represents the average of results from 3 separate experiments, and error bars represent SD, *** p < 0.001; ** p < 0.01; * p< 0.05; ns, not significant (one-way ANOVA followed by a post hoc Dunnett comparison) relative to the equivalent strain on WT THP-1 control cells. Lines indicate significance between pairs of conditions determined by Student's t test.

false