Anti-CXCR4 抗体 [EPUMBR3]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Immunohistochemical analysis of paraffin embedded Human small cell lung carcinoma tissue labeling CXCR4 using ab181020.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Immunocytochemistry/Immunofluorescence analysis of Ramos (Human Burkitt's lymphoma cell line) labeling CXCR4 with purified ab181020 at 1/500 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Intracellular Flow Cytometry analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CXCR4 with purified ab181020 at 1/200 dilution (10.23 μg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

ab181020 stained Jurkat cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab181020 at 5μg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• WB

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Western blot - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Running buffer : MOPS.

Conditions : denatured/reduced.

This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab181020 (anti-CXCR4) and ab7671 (loading ctrl), overnight at 4°C. Before imaging, antibody binding was detected using labelled goat anti-rabbit (H+L; green) and labelled goat anti-mouse (H+L; red) at 1 : 10,000 dilutions for 1hr at room temperature.

All lanes:

Western blot - Anti-CXCR4 antibody [EPUMBR3] (ab181020)

Lane 1:

CHO (chinese hamster ovary cell line) whole cell lysate (negative control) at 20 µg

Lane 2:

Jurkat whole cell at 20 µg

Lane 3:

Jurkat membrane at 20 µg

Lane 4:

Jurkat nuclear (negative control) at 20 µg

Secondary

All lanes:

Goat anti-rabbit at 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 43 kDa

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• WB

Supplier Data

Western blot - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

All lanes:

Western blot - Anti-CXCR4 antibody [EPUMBR3] (ab181020) at 1/1000 dilution

All lanes:

CXCR4 stably expressed in HEK293 cells

Predicted band size: 39 kDa

false

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Immunocytochemistry-immunofluorescence using Anti-CXCR4 antibody [EPUMBR3], ab181020. Publication image from Jiang, G. et al., 2020, Adv Sci (Weinh), 32154087. Legend direct from paper.

Patient‐derived GICs efficiently captured SLNPs via CXCR4‐stimulated macropinocytosis. a) SDF1 mimic peptides (FH27/FH29/FH38) modification enhanced the cellular uptake of DiI‐labeled SLNPs. SLNPs were prepared at the peptides : DMPC, molar ratio 1 : 100, and incubated with GICs at 37 °C for 3.5 h at the DMPC concentration of 20 µg mL−1 (n = 3). The significance of the differences was evaluated by one‐way ANOVA followed by Bonferroni test (*p < 0.05, ****p < 0.0001). b) Cellular uptake of DiI‐SLNPs and colocalization of SLNPs to CXCR4. Scale bar, 100 µm. c) The FH38‐DiI‐SLNPs showed the highest percentage of colocalization with CXCR4. d) Knockdown of CXCR4 led to a reduction in the cellular uptake of DiI‐LNPs and DiI‐SLNPs (n = 3). The significance of the differences between two groups (**p < 0.01, ****p < 0.0001) was evaluated by two‐tailed Student's t‐test. e) Qualitative analysis of the GICs uptake of LNPs and SLNPs after knocking down CXCR4. Scale bar, 100 µm. f) Colocalization of DiI‐LNPs and DiI‐SLNPs to macropinocytosis marker FITC‐70 kDa dextran in GICs in the absence/presence of EIPA (150 x 10−6m). Scale bar, 100 µm.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [EPUMBR3] (AB181020)

Immunocytochemistry-immunofluorescence using Anti-CXCR4 antibody [EPUMBR3], ab181020. Publication image from Jiang, G. et al., 2020, Adv Sci (Weinh), 32154087. Legend direct from paper.

Patient‐derived GICs efficiently captured SLNPs via CXCR4‐stimulated macropinocytosis. a) SDF1 mimic peptides (FH27/FH29/FH38) modification enhanced the cellular uptake of DiI‐labeled SLNPs. SLNPs were prepared at the peptides : DMPC, molar ratio 1 : 100, and incubated with GICs at 37 °C for 3.5 h at the DMPC concentration of 20 µg mL−1 (n = 3). The significance of the differences was evaluated by one‐way ANOVA followed by Bonferroni test (*p < 0.05, ****p < 0.0001). b) Cellular uptake of DiI‐SLNPs and colocalization of SLNPs to CXCR4. Scale bar, 100 µm. c) The FH38‐DiI‐SLNPs showed the highest percentage of colocalization with CXCR4. d) Knockdown of CXCR4 led to a reduction in the cellular uptake of DiI‐LNPs and DiI‐SLNPs (n = 3). The significance of the differences between two groups (**p < 0.01, ****p < 0.0001) was evaluated by two‐tailed Student's t‐test. e) Qualitative analysis of the GICs uptake of LNPs and SLNPs after knocking down CXCR4. Scale bar, 100 µm. f) Colocalization of DiI‐LNPs and DiI‐SLNPs to macropinocytosis marker FITC‐70 kDa dextran in GICs in the absence/presence of EIPA (150 x 10−6m). Scale bar, 100 µm.