Immunocytochemical/immunofluorescent analysis of  4% paraformaldehyde-fixed Triton X-100 permeabilized Goat skeletal muscle-derived satellite cells stained for NSE using ab53025.

After washing in PBS, the cells were incubated with Goat Anti-Rabbit IgG H&L (FITC) (ab97050) (Green).

ab84235 staining melanoma inhibitory activity in a human melanoma cell line by Flow Cytometry. Cells were harvested with EDTA and washed in PBS. Cells were pemeabilized with saponine. The sample was incubated with the primary antibody (1/100 in PBS) for 15 minutes at 20°C. A FITC-conjugated goat anti-rabbit IgG H&L (ab97050) (1/100) was used as the secondary antibody.

 

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Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

Preparation:

• Fix in 3% PFA in PBS for 30 min at RT
• Incubate in 7.5% sucrose-PBS for 3h at RT
• Incubate in 15% sucrose-PBS at 4 degree Celsius overnight
• Embed the EBs in tissue-Tek OCT compound
• Cut frozen sections to 4-20 µm thickness

 

Primary antibodies:
Mouse anti-Ki67, 1:100
Rabbit anti-laminin alpha 1 (basement marker)

Secondary antibodies:
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (AMCA) (ab47052), 1:100
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab97050), 1:100

 

The FACS staining was perform on THP-1 cell lines with Rabbit polyclonal to CCR2 (ab21667) at a 1/100 dillution for 30 min at 4C. The secondary antibody was ab97050 used at 1/100 for 20min at 4C. The buffer used was PBS/BSA (0.5%)/Azide (0.05%). No fixation or permeabilization was performed and gating was done on alive cells.

This image is courtesy of an anonymous Abreview

Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

Preparation:

• Fix in 3%PFA in PBS for 30 min at RT
• Incubate in 7.5% sucrose-PBS for 3h at RT
• Incubate in 15% sucrose-PBS at 4 degree Celsius overnight
• Embed the EBs in tissue-Tek OCT compound
• Cut frozen sections to 4-20 µm thickness

Primary antibody: Rabbit anti-laminin alpha 1, 1:400
Secondary antibody: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC) (ab97050)
F-actin was stained with CytoPainter F-actin staining kit (blue) (ab112124), 1:1000
Nuclei were counterstained stained with DRAQ7^TM (ab109202), 1:1000

The FACS staining was performed on HEK cells expressing human CCR5 with PBS/BSA (0.5%)/Azide (0.05%) as FACS buffer. We used a primary polyclonal rabbit antibody against CCR5 at a 1/100 dillution, incubated for 1 hour at 4C. The secondary antibody was ab97050 used at a concentration of 1/100 for 30min at 4C. No fixation or permeabilization was performed and gaiting was done on alive cells.

This image is courtesy of an anonymous Abreview