Anti IgG (H&L)

DyLightTM 488 conjugated anti Rabbit IgG was used to demonstrate 2 color STED immunofluorescence microscopy. Methanol fixed A431 cells were blocked with normal goat serum. The cells were then probed with 0.4 ug/mL final concentration of anti HDAC and detected with 0.2 ug/mL DyLightTM488 conjugated Anti RABBIT IgG [GOAT] secondary antibody (colored GREEN). Also shown in this 2 color STED image is Rocklands a tubulin monoclonal antibody [MOUSE] (p/n 200 301 880) detected with ATTOR 425 conjugated anti MOUSE IgG [GOAT] (610 151 121) secondary antibody (colored RED). Image courtesy of Myriam Gastard, Leica Microsystems, USA.

Properties of DyLightTM Fluorescent Dyes.

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

Western Blot of Unconjugated Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n 611-101-122). Lane M: 3 μl Molecular Ladder. Lane 1: Rabbit IgG whole molecule (p/n 011-0102). Lane 2: Rabbit IgG F(ab) Fragment (p/n 011-0105). Lane 3: Rabbit IgG F(c) Fragment (p/n 010-0103). Lane 4: Rabbit IgM Whole Molecule (p/n 011-0107). Lane 5: Normal Rabbit Serum (p/n B309). All samples were reduced. Load: 50 ng per lane. Block: MB-070 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (GOAT) Antibody (Min X Bv, Ch, Gt, GP, Ham, Hs, Hu, Ms, Rt & Sh Serum Proteins) (p/n 611-101-122) 1:1,000 for 60 min at RT. Secondary antibody: Anti-Goat IgG (DONKEY) Peroxidase Conjugated Antibody (p/n CUST10) 1:40,000 in MB-070 for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM. Rabbit F(c) migrates slightly higher.

DyLight？ 488 conjugated anti-Rabbit IgG was used to demonstrate 2 color STED immunofluorescence microscopy. Methanol fixed A431 cells were blocked with normal goat serum. The cells were then probed with 0.4 μg/mL final concentration of anti-HDAC and detected with 0.2 μg/mL DyLight？488 conjugated Anti-RABBIT IgG [GOAT] secondary antibody (colored GREEN). Also shown in this 2-color STED image is Rockland’s a-tubulin monoclonal antibody [MOUSE] (p/n 200-301-880) detected with ATTOR 425 conjugated anti-MOUSE IgG [GOAT] (610-151-121) secondary antibody (colored RED). Image courtesy of Myriam Gastard, Leica Microsystems, USA.