Anti-Profilin 1 antibody [EPR6303]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6303] (AB133529)

Immunohistochemistry analysis of Paraffin Embedded Human kidney tissue labelling Profilin 1 with ab133529 at 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Profilin 1 antibody [EPR6303] (AB133529)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133529 observed at 15 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab133529 was shown to specifically react with in wild-type HAP1 cells as signal was lost in PFN1 knockout cells. Wild-type and PFN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. ab133529 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Profilin 1 antibody [EPR6303] (ab133529) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

PFN1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

Jurkat whole cell lysate at 20 µg

Predicted band size: 15 kDa

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• WB

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Western blot - Anti-Profilin 1 antibody [EPR6303] (AB133529)

All lanes:

Western blot - Anti-Profilin 1 antibody [EPR6303] (ab133529) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

JAR cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Predicted band size: 15 kDa

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• WB

Lab

Western blot - Anti-Profilin 1 antibody [EPR6303] (AB133529)

ab133529 was shown to react with PFN1 in wild-type HAP1 cells in Western blot with loss of signal observed in a PFN1 knockout cell line. Wild-type HAP1 and PFN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab133529 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Profilin 1 antibody [EPR6303] (ab133529) at 1/10000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

PFN1 knock-out HAP1 lysate at 20 µg

Observed band size: 15 kDa

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• WB

CiteAb

Western blot - Anti-Profilin 1 antibody [EPR6303] (AB133529)

Profilin 1 western blot using anti-Profilin 1 antibody [EPR6303] ab133529. Publication image and figure legend from Shen, K., Xi, Z., et al., 2016, Oncotarget, PubMed 27494863.

ab133529 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 ab133529 please see the product overview.

PFN 1 mediates GUTK action on HCC cell motility(A) Representative silver-stained-2-DE images of HepG2 cells treated with or without GUTK (20 μM) for 24 h. Differentially expressed spots are shown by the arrows. (B) Ingenuity pathway analysis (Ingenuity Systems, www.Ingenuity.com). Bars represent molecular and cellular functions that are significantly changed following incubation with GUTK. (C) Cropped and enlarged regions of the differently expressed PFN1 spot (top), and PFN1 protein expression in HepG2 cells treated with GUTK for 24 h by western blot (bottom). (D) PFN1 in HepG2 cells transiently transfected with PFN1 or control vector for 24–72 h by western blot. GAPDH served as a loading control. (E, F) The migration and invasion were determined in HepG2 cells transiently transfected with PFN1 or control vector for 24 h and 48 h. Data are shown as mean ± SEM; ***p < 0.001 vs. cells transfected with control vector. n = 3. (G) PFN1 in HepG2 cells transiently transfected with two individual PFN1 siRNAs (si.PFN1-1 and si.PFN1-2) or control siRNA (si.Control) for 24–72 h by western blot. (H, I) The migration and invasion were determined in HepG2 cells transiently transfected with PFN1 siRNAs or control siRNA for 24 h and 48 h. Data are shown as mean ± SEM; **p < 0.01, ***p < 0.001 vs. cells transfected with si.Control. n = 3.

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