Cytospins of mouse pre-B cells and cryopreserved Epstein-Barr virus (EBV)-transformed lymphocytes from members of the index family, were fixed with 4% paraformaldehyde and washed with phosphate-buffered saline (PBS), followed by a 30 min incubation in a blocking/permeabilization solution of 10% normal goat serum (NGS)/0.1% Triton-X 100/PBS, and then incubated for 1 hr with primary anti-IKAROS antibody (H-100, Santa Cruz Biotechnology, Santa Cruz, CA) diluted in 3% NGS/0.1% Triton-X 100/PBS. Slides were washed three times in PBS, and then incubated for 45 min in 3% NGS/0.1% Triton-X 100/PBS containing a secondary antibody conjugated to Ig-Alexa Fluor 488 (Invitrogen, Carlsbad, CA). Slides were washed three times in PBS and mounted with Vectashield (Vector Labs, Burlingame, CA) containing 4′-6-diamidino-2-phenylindol (DAPI). All steps were carried out at room temperature. Images were captured using a Nikon C2 confocal fluorescence microscope and analyzed for nuclear versus cytoplasmic localization using NIS Elements software.
Ig alexa fluor 488
Manufactured by Thermo Fisher Scientific
Ig-Alexa Fluor 488 is a fluorescent labeling reagent used for the detection and visualization of antibodies or other proteins in various biological applications. It consists of an antibody-binding molecule conjugated to the Alexa Fluor 488 fluorescent dye. The Alexa Fluor 488 dye provides bright green fluorescence upon excitation with a suitable light source.
Automatically generated - may contain errors
Lab products found in correlation
Vectashield, by Vector Laboratories (1 mentions)
C2 confocal fluorescence microscope, by Nikon (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
γh2ax, by Merck Group (1 mentions)
Ix51 fluorescence microscope, by Olympus (1 mentions)
Anti mbrd7 polyclonal antibody, by Proteintech (1 mentions)
2 protocols using ig alexa fluor 488
1
Immunofluorescence Localization of IKAROS
2
Immunohistochemical Analysis of Testis
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The preparation of testis slides was processed as previously described in IHC protocol. Antibodies were used as followed: anti-mBRD7 polyclonal antibody (ProteinTech, 1:1000), γH2AX (Millipore 05–636, 1∶1000). Slides were washed three times in PBS, and then incubated for 1 hour at room temperature in 3% NGS, 0.1% Triton-X 100 in PBS containing the relevant secondary antibodies conjugated to Ig-Alexa Fluor 594 or Ig-Alexa Fluor 488 (1∶500 dilutions; Invitrogen). Slides were washed three times in PBS before incubate with DAPI (1∶1000 dilutions; Invitrogen). Images of tissue sections were photographed using a Olympus IX51 fluorescence microscope.
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