5×104 MSCs were lysed on ice with RIPA lysate buffer supplemented with protease inhibitor cocktail (Roche). Total protein concentration was determined using the BCA protein assay kit (Pierce). Antibodies used: goat anti-MIF (1:250, Santa Cruz), rabbit anti-CXCR4 (1:500, Boster Biological Technology), rabbit anti-CXCR2 (1:500, Ab Frontier), mouse anti-CD74 (1:100, Santa Cruz) rabbit phospho p44/42 (ERK1/2) (Thr202/Tyr204) (1:1000, Cell signaling), rabbit phospho SAPK/JNK (Thr183/Tyr185) (1:1000, Cell signaling), goat anti-p44/42 (ERK1/2) (1:5000, Santa Cruz), rabbit SAPK/JNK (1:1000, Cell signaling). Secondary antibodies used: anti-goat HRP, anti-rabbit HRP or anti-mouse HRP (1:7500, Sigma). Membranes were incubated with ECL substrate (Millipore), exposed and developed following manufacturer’s recommendations.
Mouse anti cd74
Manufactured by Santa Cruz Biotechnology
Mouse anti-CD74 is a primary antibody that binds to the CD74 antigen. CD74 is a type II transmembrane glycoprotein expressed on the surface of antigen-presenting cells, including B cells, monocytes, macrophages, and dendritic cells. The antibody can be used for the detection and analysis of CD74 expression in various applications.
Automatically generated - may contain errors
2 protocols using mouse anti cd74
1
Quantification of MIF and CXCR Signaling in MSCs
2
Colocalization of CXCR4 and CD74 in Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Endothelial cells were cultured on cover slips in 24-well plates at
a concentration 4 × 104 cells per well for 24 h at
37 °C. Next, the cells were infected with P.
gingivalis at an MOI of 100 for 24 h. The cells were subsequently
washed and fixed, after which they were incubated in 1% BSA in 0.1% PBS-Tween
for 1 h. Afterwards, the samples were incubated with mouse anti-CD74 (1:100;
Santa Cruz) and rabbit anti-CXCR4 (1:200; Abcam) overnight at 4 °C. Donkey
anti-mouse IgG labeled with PE (1:20; Proteintech) and goat anti-rabbit IgG
labeled with FITC (1:20; Proteintech) were used as the secondary antibodies and
were incubated with the cells for 1 h at room temperature. DAPI (Boster, Wuhan,
China) was used to stain the cell nucleus (pseudo-colored blue), and the cells
were observed using a fluorescence microscope (Nikon 80i, Japan) to evaluate the
colocalization between CXCR4 and CD74.
a concentration 4 × 104 cells per well for 24 h at
37 °C. Next, the cells were infected with P.
gingivalis at an MOI of 100 for 24 h. The cells were subsequently
washed and fixed, after which they were incubated in 1% BSA in 0.1% PBS-Tween
for 1 h. Afterwards, the samples were incubated with mouse anti-CD74 (1:100;
Santa Cruz) and rabbit anti-CXCR4 (1:200; Abcam) overnight at 4 °C. Donkey
anti-mouse IgG labeled with PE (1:20; Proteintech) and goat anti-rabbit IgG
labeled with FITC (1:20; Proteintech) were used as the secondary antibodies and
were incubated with the cells for 1 h at room temperature. DAPI (Boster, Wuhan,
China) was used to stain the cell nucleus (pseudo-colored blue), and the cells
were observed using a fluorescence microscope (Nikon 80i, Japan) to evaluate the
colocalization between CXCR4 and CD74.
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