Western blot - CXCR4 antibody (ab2074)

Anti-CXCR4 antibody (ab2074) at 1/2000 dilution + HeLa whole cell lysate

Predicted band size : 39 kDa

Immunocytochemistry - CXCR4 antibody (ab2074)

Immunocytochemistry of CXCR4 in HeLa cells with CXCR4 antibody ab2074 at 2 µg/ml.

Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)

ab2074 at 1/100 staining differentiated rat skeletal myocytes by ICC/IF. The cells were 2% paraformaldehyde fixed for 15 minutes and then incubated cells with ab2074 overnight at 4°C. The image demonstrates CXCR4 expressing skeletal myocytes (green-cytoplasmic and cell membrane localization) with DAPI (blue-nuclear counter stain) [Right panel] and DIC (phase) image of the same skeletal myocytes ([left panel].

This image is courtesy of an Abreview submitted by Dr Mal Niladri

Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)

ab2074 staining CXCR4 expressing cells (of mesenchymal origin) by ICC/IF. The cells were methanol fixed and blocked with BSA prior to incubation with the antibody for 1 hour. A FITC conjugated swine anti-rabbit antibody was used as the secondary. Nuclei were counterstained with DAPI.

This image is courtesy of an anonymous Abreview

Flow Cytometry - CXCR4 antibody (ab2074)

ab2074 staining cells from human B cell lymphoma by flow cytometry.  Cells were cultured and prepared in FACS buffer.  The primary antibody was diluted 1/50 and incubated with the sample for 20 minutes at 4°C. A FITC conjugated swine anti-rabbit antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

Western blot - CXCR4 antibody (ab2074)

Lane 1 : Anti-CXCR4 antibody (ab2074) at 1/250 dilution
Lane 2 : Anti-CXCR4 antibody (ab2074) at 1/500 dilution
Lane 3 : Anti-CXCR4 antibody (ab2074) at 1/750 dilution
Lane 4 : Anti-CXCR4 antibody (ab2074) at 1/1000 dilution
Lane 5 : Anti-CXCR4 antibody (ab2074) at 1/1500 dilution
Lane 6 : Anti-CXCR4 antibody (ab2074) at 1/2000 dilution

Lane 1 : Lysate prepared from human Ewing`s sarcoma cells
Lane 2 : Lysate prepared from human Ewing`s sarcoma cells
Lane 3 : Lysate prepared from human Ewing`s sarcoma cells
Lane 4 : Lysate prepared from human Ewing`s sarcoma cells
Lane 5 : Lysate prepared from human Ewing`s sarcoma cells
Lane 6 : Lysate prepared from human Ewing`s sarcoma cells

Lysates/proteins at 500000 cells per lane.

Secondary
HRP-conjugated goat polyclonal to rabbit IgG at 1/4000 dilution

Performed under reducing conditions.

Predicted band size : 39 kDa
Observed band size : 43 kDa (why is the actual band size different from the predicted?)


Exposure time : 5 minutes

This image is a courtesy of Sabine Topka

Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)

ab2074 staining CXCR4 in rat bone marrow cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed, permeabilised in 0.1% Triton X-100 and then blocked using 5% serum for 1 hour at 25ºC. Samples were then incubated with primary antibody at 1/100 for 12 hours at 4ºC. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/250 dilution. DAPI was used to stain the cell nuclei (blue).

Image courtesy of an anonymous Abreview

Immunohistochemistry (Frozen sections) - CXCR4 antibody (ab2074)

ab2074 staining CXCR4 in rat kidney tubular epithelium tissue by Immunohistochemistry (Frozen sections). The tissue was formaldehyde fixed and then blocked using 2% BSA for 2 hours at 25ºC. Samples were then incubated with primary antibody at 1/1000 for 9 hours at 4ºC. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).

Image courtesy of an anonymous Abreview.

Immunocytochemistry/ Immunofluorescence - CXCR4 antibody (ab2074)

ICC/IF image of ab2074 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2074, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.