Human monocytes were treated with 200 ng/ml CCL2 or an equal volume of 0.1% BSA in PBS, the CCL2 diluent, for 15 minutes. Cells were then fixed with 2 % paraformaldehyde in PBS for 1 hour, permeabilizated with 0.01% triton, and incubated in blocking solution containing 0.5 M EDTA, 1% fish gelatin, 1% Ig-free BSA, 1% horse serum and 1% human serum for 30 minutes. After blocking, cells were stained with anti-tubulin antibody (1:10.000, #T9026, Sigma) for 1 hour at room temperature. The cells were washed and then incubated with the secondary antibody, anti-mouse IgG F(ab′)2 fragment-FITC (1:100, #F2883, Sigma) mixed with Texas Red®-X phalloidin ( 15ul/ml, #T7471, Invitrogen, Grand Island, NY) for 2 hours at room temperature. Samples were washed and then mounted in Prolong Gold antifade reagent (P36931, Invitrogen) and examined by confocal microscopy using a Leica SP2 confocal microscope.
Anti mouse igg f ab 2 fragment fitc
Manufactured by Merck Group
Sourced in United States
Anti-mouse IgG F(ab′)2 fragment-FITC is a laboratory reagent that consists of the F(ab′)2 fragment of mouse immunoglobulin G (IgG) conjugated to the fluorescent dye Fluorescein Isothiocyanate (FITC). It is used for various applications in biomedical research and diagnostics.
Automatically generated - may contain errors
Lab products found in correlation
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Sp2 confocal microscope, by Leica (1 mentions)
Anti tubulin antibody, by Merck Group (1 mentions)
Texas red x phalloidin, by Thermo Fisher Scientific (1 mentions)
Facscalibur, by BD (1 mentions)
Guava easycyte ht flow cytometer, by Merck Group (1 mentions)
Countess cell counter, by Thermo Fisher Scientific (1 mentions)
Kaluza analysis 1, by Beckman Coulter (1 mentions)
2 protocols using anti mouse igg f ab 2 fragment fitc
1
Visualization of Cytoskeleton Dynamics
2
Flow Cytometric Analysis of Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The OS cell lines were harvested and cell numbers were determined by utilizing the Countess Cell Counter (Invitrogen) followed by dilution in flow buffer (Dulbecco’s PBS with 0.5% BSA and 0.1% NaN3). The cell suspension was distributed in a 96 well plate with 2 × 105 cells per sample well. Primary antibodies were added in a concentration of 10 μg/ml and cells were incubated at 4°C for 30 min before three washes in 200 μl flow buffer. Secondary antibody, anti-mouse IgG F(ab’)2 fragment-FITC (Sigma-Aldrich, St Louis, MO, USA) or anti-human IgG-FITC (US Biologicals, Salem, MA, USA), was added and incubated for 30 min and washed as in the previous step. All wash steps were performed by centrifugation at 1,200 rpm for 5 min.
Washed cell pellets were dissolved in flow buffer and analyzed in a FACS Calibur (BD Bioscience, Franklin Lakes, NJ, USA) or Guava EasyCyte HT flow cytometer (Merck Millipore, Darmstadt, Germany). The flow analysis was replicated and reproduced at least three times for all cell lines. The data were analyzed with Kaluza Analysis 1.3 software (Beckman Coulter, Brea, CA, USA).
Washed cell pellets were dissolved in flow buffer and analyzed in a FACS Calibur (BD Bioscience, Franklin Lakes, NJ, USA) or Guava EasyCyte HT flow cytometer (Merck Millipore, Darmstadt, Germany). The flow analysis was replicated and reproduced at least three times for all cell lines. The data were analyzed with Kaluza Analysis 1.3 software (Beckman Coulter, Brea, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!