4-Color Imaging with DyLight 405 Conjugates
Our DyLight 405-conjugated secondary antibodies are excited maximally at about 400 nm and fluoresce with a peak at about 421 nm. They are very bright and photostable when used with microscopes or flow cytometers with a 405 nm laser and a 420 nm emission filter (Figure 2).
They will not work well with a 450 nm emission filter.
Under these conditions, it is possible to perform effective 4-color imaging with good color separation, good photostability, and high sensitivity in both aqueous and permanent mounting media. The combination of DyLight 405, Alexa Fluor 488, Rhodamine Red-X, and Alexa Fluor 647 provides for maximum color separation (see Spectra Viewer).
Other 4-color dye combinations, which may be equally effective but have slightly less color separation, include DyLight 405, Alexa Fluor 488, Cy3, and Alexa Fluor 647.
Figure 1. Mouse retinal tissue stained with 4 colors, including DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO). Sections were mounted in n-propyl gallate-glycerol for imaging with an Olympus Fluoview 1000 Laser Scanning confocal microscope equipped with a 405 nm laser and a krypton-argon laser. Images are courtesy of Gabe Luna, Neuroscience Research Institute, UC Santa Barbara.
Figure 2. Mouse retinal tissue stained with 4 colors including DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO). Sections were mounted in n-propyl gallate-glycerol for imaging with an Olympus Fluoview 1000 Laser Scanning confocal microscope equipped with a 405 nm laser and a krypton-argon laser. Images are courtesy of Gabe Luna, Neuroscience Research Institute, UC Santa Barbara.
The following table lists highly adsorbed affinity-purified secondary antibodies, streptavidin, and immunoglobulin controls conjugated with DyLight 405 as offered by Jackson ImmunoResearch for use only in 4-color labeling protocols using a 405 nm laser-equipped confocal microscope. For a complete listing of other DyLight 405 conjugates for 1-, 2-, and 3-color labeling, see complete tables of
Whole IgG Secondary Antibodies and F(ab')2 Fragment Secondary Antibodies.
|
Product Description
|
DyLight
405
A=400, E=421
|
|
Streptavidin
|
|
Donkey Anti-Chicken IgY
†(IgG)(H+L) ML*
(min X Bov, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Goat IgG (H+L)
♦
ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
|
|
Donkey Anti-Guinea Pig IgG (H+L)
ML*
(min X Bov, Ck, Gt, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Human IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Mouse IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
|
|
Donkey Anti-Mouse IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb,
Rat Shp Sr Prot)**
|
|
Goat Anti-Mouse IgG (H+L)
ML*
(min X Hu, Bov, Hrs, Rb, Sw Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2a Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2b Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2c Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG (H+L)
ML*
(min X
Rat, Hu, Bov,Hrs, Rb Sr Prot)
|
|
Donkey Anti-Rabbit IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rat, Shp Sr Prot)
|
|
Goat Anti-Rabbit IgG (H+L)
ML*
(min X Hu, Ms, Rat Sr Prot)
|
|
Donkey Anti-Rat IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
|
|
Donkey Anti-Rat IgG (H+L)
** ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu,
Ms, Rb, Shp Sr Prot)
|
|
Goat Anti-Rat IgG (H+L)
ML*
(min X
Ms, Hu, Bov, Hrs, Rb Sr Prot)
|
|
Donkey Anti-Sheep IgG (H+L)
♦
ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
|
|
|
ChromPure Donkey IgG, whole molecule
|
|
|
ChromPure Goat IgG, whole molecule
|
|
|
*
|
ML=Multiple Labeling (see Multiple Labeling for an explanation.).
|
|
|
†
|
IgY is the original designation for the IgG-like protein found in both serum and egg yolk.
|
|
♦
|
Warning: BSA and dry milk may contain IgG which will react with this antibody. Use of BSA and or dry milk to block or dilute this antibody may increase background and or reduce secondary antibody titer.
|
|
**
|
Caution: See Selection of Affinity-Purified Antibodies before selecting an antibody adsorbed against closely related species.4-Color Imaging with DyLight 405 Conjugates
Our DyLight 405-conjugated secondary antibodies are excited maximally at about 400 nm and fluoresce with a peak at about 421 nm. They are very bright and photostable when used with microscopes or flow cytometers with a 405 nm laser and a 420 nm emission filter (Figure 2).
They will not work well with a 450 nm emission filter.
Under these conditions, it is possible to perform effective 4-color imaging with good color separation, good photostability, and high sensitivity in both aqueous and permanent mounting media. The combination of DyLight 405, Alexa Fluor 488, Rhodamine Red-X, and Alexa Fluor 647 provides for maximum color separation (see Spectra Viewer).
Other 4-color dye combinations, which may be equally effective but have slightly less color separation, include DyLight 405, Alexa Fluor 488, Cy3, and Alexa Fluor 647.
Figure 1. Mouse retinal tissue stained with 4 colors, including DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO). Sections were mounted in n-propyl gallate-glycerol for imaging with an Olympus Fluoview 1000 Laser Scanning confocal microscope equipped with a 405 nm laser and a krypton-argon laser. Images are courtesy of Gabe Luna, Neuroscience Research Institute, UC Santa Barbara.
Figure 2. Mouse retinal tissue stained with 4 colors including DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO). Sections were mounted in n-propyl gallate-glycerol for imaging with an Olympus Fluoview 1000 Laser Scanning confocal microscope equipped with a 405 nm laser and a krypton-argon laser. Images are courtesy of Gabe Luna, Neuroscience Research Institute, UC Santa Barbara.
The following table lists highly adsorbed affinity-purified secondary antibodies, streptavidin, and immunoglobulin controls conjugated with DyLight 405 as offered by Jackson ImmunoResearch for use only in 4-color labeling protocols using a 405 nm laser-equipped confocal microscope. For a complete listing of other DyLight 405 conjugates for 1-, 2-, and 3-color labeling, see complete tables of
Whole IgG Secondary Antibodies and F(ab')2 Fragment Secondary Antibodies.
|
Product Description
|
DyLight
405
A=400, E=421
|
|
Streptavidin
|
|
Donkey Anti-Chicken IgY
†(IgG)(H+L) ML*
(min X Bov, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Goat IgG (H+L)
♦
ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
|
|
Donkey Anti-Guinea Pig IgG (H+L)
ML*
(min X Bov, Ck, Gt, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Human IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Ms, Rb, Rat, Shp Sr Prot)
|
|
Donkey Anti-Mouse IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
|
|
Donkey Anti-Mouse IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb,
Rat Shp Sr Prot)**
|
|
Goat Anti-Mouse IgG (H+L)
ML*
(min X Hu, Bov, Hrs, Rb, Sw Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2a Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2b Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 2c Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific
ML*
(min X Hu, Bov, Rb Sr Prot)
|
|
Goat Anti-Mouse IgG (H+L)
ML*
(min X
Rat, Hu, Bov,Hrs, Rb Sr Prot)
|
|
Donkey Anti-Rabbit IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rat, Shp Sr Prot)
|
|
Goat Anti-Rabbit IgG (H+L)
ML*
(min X Hu, Ms, Rat Sr Prot)
|
|
Donkey Anti-Rat IgG (H+L)
ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
|
|
Donkey Anti-Rat IgG (H+L)
** ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu,
Ms, Rb, Shp Sr Prot)
|
|
Goat Anti-Rat IgG (H+L)
ML*
(min X
Ms, Hu, Bov, Hrs, Rb Sr Prot)
|
|
Donkey Anti-Sheep IgG (H+L)
♦
ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
|
|
|
ChromPure Donkey IgG, whole molecule
|
|
|
ChromPure Goat IgG, whole molecule
|
|
|
*
|
ML=Multiple Labeling (see Multiple Labeling for an explanation.).
|
|
|
†
|
IgY is the original designation for the IgG-like protein found in both serum and egg yolk.
|
|
♦
|
Warning: BSA and dry milk may contain IgG which will react with this antibody. Use of BSA and or dry milk to block or dilute this antibody may increase background and or reduce secondary antibody titer.
|
|
**
|
Caution: See Selection of Affinity-Purified Antibodies before selecting an antibody adsorbed against closely related species. | |
|
