Anti-CX3CR1 antibody

• WB

Lab

Western blot - Anti-CX3CR1 antibody (AB8020)

False colour image of Western blot : Anti-CX3CR1 antibody staining at 1 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab8020 was shown to bind specifically to CX3CR1. A band was observed at 42 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CX3CR1 knockout cell line ab273713 (knockout cell lysate ab275510). To generate this image, wild-type and CX3CR1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) 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-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CX3CR1 antibody (ab8020) at 1 µg/mL

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human CX3CR1 knockout THP-1 cell lysate (<a href='/en-us/products/cell-lysates/human-cx3cr1-knockout-thp-1-cell-lysate-ab275510'>ab275510</a>) at 20 µg

Lane 3:

Human Spleen cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Lane 5:

Caco-2 cell lysate at 20 µg

Predicted band size: 40 kDa

Observed band size: 42 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody (AB8020)

ab8020 at 10μg/ml staining CX3CR1 in human spleen cells by IHC

• WB

Unknown

Western blot - Anti-CX3CR1 antibody (AB8020)

All lanes:

Western blot - Anti-CX3CR1 antibody (ab8020) at 1/500 dilution

Lane 1:

THP-1 cell lysate with absence of blocking peptide

Lane 2:

THP-1 cell lysate with presence of blocking peptide

Predicted band size: 40 kDa

Observed band size: 50 kDa

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• WB

CiteAb

Western blot - Anti-CX3CR1 antibody (AB8020)

CX3CR1 western blot using anti-CX3CR1 antibody ab8020. Publication image and figure legend from Rooper, L., Gurler, H., et al., 2013, J Ovarian Res, PubMed 23958497.

ab8020 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 ab8020 please see the product overview.

CX3CR1 is required for keratinocytes differentiation. (A) Expression of CX3CR1 in parental PA-1 cell line and that stably transfected with either CX3CR1 or scrambled shRNA constructs was examined with Western blot. Actin served as a loading control. CX3CR1 expression levels were quantified using digital densitometry and normalized to the levels of actin expression. (B) Expression of cytokeratin 14 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with immunocytochemistry. CK14 in PA-1 was probed with anti-CK14 and Alexa555-conjugated anti-rabbit antibodies, and DNA was stained with 4',6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI); CK14 – red, DNA – blue. Images were taken using 5× magnification on the objective. The histogram demonstrates the average intensity of CK14 signal across the field as determined by the Zeiss AxioVision software. Data is an average of three independent experiments. *p < 0.05. (C) Expression of cytokeratins 14 and 18 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with Western blot. β-Tubulin served as a loading control. The histogram shows CK14 and CK18 expression levels. Expression of CK18 in CX3CR1shRNA-transfected PA-1 cells was arbitrarily set as 1 and expression of both CK14 and CK18 in other conditions was calculated accordingly. The data represent a typical Western blot image, and quantitative analysis was performed using three independently performed experiments. *p < 0.05. (D) Expression of CX3CL1 in SKOV-3 and PA-1 cell lines was examined with Western blot. Actin served as a loading control. SKOV-3 cell line was used as a positive control.

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