Jackson Immuno Research Inc.
specializing in secondary antibodies and conjugates

Find By Code Number

Product Filter

Whole IgG Affinity-Purified Secondary Antibodies

"Without question, I can always turn to Jackson ImmunoResearch secondary antibodies for ECL detection of any primary antibody! They produce great signal at low dilutions, last for years even through bad laboratory practice like repeatedly using the same tube every time, and come in a variety of species flavors."

W Thompson, OSHU

Leave a Review

Spotlight on...

Fluorescent Dyes

Brilliant Violet™
421 & 480

  • Combine with AF488, RR-X, & AF647 for 5-color imaging
  • Compatible with commonly used filter sets
  • Switch nuclear stain from DAPI to DRAQ5™ for additional labeling options

Learn More

Meet JIR...

Exhibit Schedule

14th Feb - Life Science Exhibits, Tufts University School of Medicine - Medford, MA

16th Feb - Life Science Exhibits, Harvard University Medical School - Boston, MA

23rd-25th Feb - ASCO-SITC Clinical Immuno-Oncology Symposium - Orlando, FL

More info

JIR tweets...

4-Color Imaging

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 (Figure 1). 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.

Figure 1. Excitation (violet) and emission (blue) spectra for DyLight 405 conjugated to an affinity-purified secondary antibody from Jackson ImmunoResearch. Quantitative comparisons should not be made since peak heights have been normalized. Spectra were obtained with an M-Series spectrofluorometer system from Photon Technology International, Inc.
figure 2 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.

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 (Figure 3).

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 3 Figure 3. Emission spectra of DyLight 405 (blue), Alexa Fluor 488 (green), Rhodamine Red-X (red), and Alexa Fluor® 647 (brown). This figure illustrates the relative shape and position of each fluorophore emission peak following conjugation to antibodies. It shows that effective 4-color imaging can be performed with maximum color separation using these dyes. Quantitative comparisons should not be made since peak heights have been normalized. All spectra were obtained with an M-Series spectrofluorometer system from Photon Technology International, Inc.
figure 4 Figure 4. 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 other tables in this Web site.

Product Description
DyLight
405
A=400, E=421
Streptavidin
$162 / 1.0 mg
016-470-084
Donkey Anti-Chicken IgY (IgG)(H+L) ML*
(min X Bov, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
130 / 0.5 mg
703-475-155
Donkey Anti-Goat IgG (H+L) ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
130 / 0.5 mg
705-475-147
Donkey Anti-Guinea Pig IgG (H+L) ML*
(min X Bov, Ck, Gt, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot)
130 / 0.5 mg
706-475-148
Donkey Anti-Human IgG (H+L) ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Ms, Rb, Rat, Shp Sr Prot)
130 / 0.5 mg
709-475-149
Donkey Anti-Mouse IgG (H+L) ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
130 / 0.5 mg
715-475-150
Donkey Anti-Mouse IgG (H+L) ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Rat Shp Sr Prot)**
184 / 0.5 mg
715-475-151
Goat Anti-Mouse IgG (H+L) ML*
(min X Hu, Bov, Hrs, Rb, Sw Sr Prot)
142 / 1.0
115-475-146
Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific ML*
(min X Hu, Bov, Rb Sr Prot)
230 / 0.5 mg
115-475-205
Goat Anti-Mouse IgG, Fcγ Subclass 2a Specific ML*
(min X Hu, Bov, Rb Sr Prot)
230 / 0.5 mg
115-475-206
Goat Anti-Mouse IgG, Fcγ Subclass 2b Specific ML*
(min X Hu, Bov, Rb Sr Prot)
230 /0.5 mg
115-475-207
Goat Anti-Mouse IgG, Fcγ Subclass 2c Specific ML*
(min X Hu, Bov, Rb Sr Prot)
230 / 0.5 mg
115-475-208
Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific ML*
(min X Hu, Bov, Rb Sr Prot)
230 / 0.5 mg
115-475-209
Goat Anti-Mouse IgG (H+L) ML*
(min X Rat, Hu, Bov,Hrs, Rb Sr Prot)
184 / 0.5 mg
115-475-166
Donkey Anti-Rabbit IgG (H+L) ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rat, Shp Sr Prot)
130 / 0.5 mg
711-475-152
Goat Anti-Rabbit IgG (H+L) ML*
(min X Hu, Ms, Rat Sr Prot)
142 / 1.0 mg
111-475-144
Donkey Anti-Rat IgG (H+L) ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Rb, Shp Sr Prot)
130 / 0.5 mg
712-475-150
Donkey Anti-Rat IgG (H+L) **                              ML*
(min X Bov, Ck, Gt, GP, Sy Hms, Hrs, Hu, Ms, Rb, Shp Sr Prot)
184 / 0.5 mg
712-475-153
Goat Anti-Rat IgG (H+L) ML*
(min X Ms, Hu, Bov, Hrs, Rb Sr Prot)
184 / 0.5 mg
112-475-167
Donkey Anti-Sheep IgG (H+L) ML*
(min X Ck, GP, Sy Hms, Hrs, Hu, Ms, Rb, Rat Sr Prot)
130 / 0.5 mg
713-475-147
ChromPure Donkey IgG, whole molecule
55 / 1.0 mg
017-470-003
ChromPure Goat IgG, whole molecule
55 / 1.0 mg
005-470-003
*

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.