Anti IgG (H&L)

Rockland DyLight and ATTO dye conjugated antibodies provide high signal and low background for confocal microscopy and high resolution Stimulated Emission Depletion (STED) Microscopy. Both Dylight and Atto conjugated secondary antibodies maintained robust, intense signal during repeated laser excitation and de-excitation used during STED microscopy. Shown here are:
A. (Green) Mouse anti NuP (NuP=Nuclear Pore Protein) detected with Dylight 488 Goat anti mouse (610-141-121)
B. (Red) Rabbit Anti Ezh1/2 Pab (Ezh=enhancer of zeste homology) with detection by Rockland ATTO 425 conjugated Goat anti Rabbit (611-151-122)
C. (Red and Green) Images combined.
Data was collected on a STED-CW TCS-SP5 Confocal system (Leica Microsystems) equipped with a DFC 350FX Camera allowing sequential acquisition in wide-field, confocal and STED CW imaging modes and provided courtesy of:
Myriam Gastard, PhD, personal communication, Leica Microsystems, Inc. USA

Western Blot showing detection of Mouse IgG, heavy and light chain. 100 ng of Mouse IgG (Lane 2) was run on a 4-20% gel and transferred to 0.45 μm nitrocellulose. After blocking with 1% BSA-TTBS (p/n MB-013, diluted to 1X) 30 min at 20C, Anti-MOUSE IgG (H&L) (GOAT) Antibody DyLight^TM 488 Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Rb Rt & Sh Serum Proteins) (p/n 610-141-121) secondary antibody was used at 1:1000 in Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) and imaged using the Bio-Rad VersaDoc^(R) 4000 MP. Molecular weight markers are in lane 1.

DyLight^TM dyes can be used for multi-color immunofluorescence microscopy with uniform fluorescence intensity throughout the image. DyLight^TM dyes are exceptionally bright and photostable and are optimized for microscopy and microarray detection methods. This image shows anti-histone detection using a DyLight^TM 488 conjugate (green). Anti-Tubulin was detected using a DyLight^TM 549 conjugate (red). Nuclei were counter-stained using DAPI (blue). The image was captured using an Axio Imager.Z1 (Zeiss Micro Imaging Inc).

Rockland DyLight and ATTO dye conjugated antibodies provide high signal and low background for confocal microscopy (upper images) and high resolution Stimulated Emission Depletion (STED) Microscopy (lower images). Both Dylight and ATTO conjugated secondary antibodies maintained robust, intense signal during repeated laser excitation and de-excitation used during STED microscopy. Shown here are:
A. (Green) Mouse anti NuP (NuP=Nuclear Pore Protein) detected with Dylight 488 Goat anti mouse (610-141-121)
B. (Red) Rabbit Anti Ezh1/2 Pab (Ezh=enhancer of zeste homology) with detection by Rockland ATTO 425 conjugated Goat anti Rabbit (611-151-122)
(Red and Green) Images combined.
Data was collected on a STED-CW TCS-SP5 Confocal system (Leica Microsystems) equipped with a DFC 350FX Camera allowing sequential acquisition in widefield, confocal and STED CW imaging modes and provided courtesy of:
Myriam Gastard, PhD, personal communication, Leica Microsystems, Inc. USA

Properties of DyLight^TM Fluorescent Dyes.

Rockland Dylight 488 Goat Anti Mouse IgG antibody-Immunofluorescence
Cell Type: A431 cells
Fixation: 4% paraformaldehyde 10 min
Permeablization: 0.5% Triton X 30 min
Primary Ab: 200-301-880 lot 28977 1:250 72 hours 4C
Secondary Ab: 610-141-121 lot 21286 1:1000 overnight 4C