Anti-CXCR4 (phospho S324 + S325) antibody [RP23040164]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 (phospho S324 + S325) Antibody [RP23040164] (AB313465)

For immunofluorescence analysis of Jurkat cells fixed and permeabilized for detection of endogenous CXCR4 [pS324/325] using ab313465 (5 ug/ml) and labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate ( 1 : 2000). Nuclei (blue) were stained using with DAPI , and Rhodamine Phalloidin ( 1 : 300) was used for cytoskeletal F-actin (red) staining. Detection and localization of CXCR4 [pS324/325] (green)on the membrane can be clearly observed in cells treated with PMA (40 ug/ml, 30 min) as compared to untreated cells. Antibody specificity was demonstrated by competition with the CXCR4 [pS324/325] phospho peptide, which results in loss of signal. No competition was observed with the non-phospho peptide. The images were captured at 60X magnification.

• WB

Supplier Data

Western blot - Anti-CXCR4 (phospho S324 + S325) Antibody [RP23040164] (AB313465)

A 55 kDa band corresponding to CXCR4 [pS324/425] was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using 4-12% Bis-Tris gel. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk.

All lanes:

Western blot - Anti-CXCR4 (phospho S324 + S325) Antibody [RP23040164] (ab313465) at 2.5 µg/mL

Lane 1:

Membrane enriched extracts of HEK-293 at 30 µg

Lane 2:

Membrane enriched extracts of A549 at 30 µg

Lane 3:

Membrane enriched extracts of MCF-7 at 30 µg

Lane 4:

Membrane enriched extracts of LNCaP at 30 µg

Lane 5:

Membrane enriched extracts of HeLa at 30 µg

Secondary

All lanes:

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

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