StarBright™ Blue 700 Fluorescent Secondary Antibodies

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StarBright Fluorescent Secondary Antibodies

Overview

Triplex Blot Example
Triplex blot image
Simultaneous detection of multiple proteins (ATG7, AKR1C2, and tubulin) on a single blot using fluorescent antibodies.
Target Protein Color 1º Antibody 2º Antibody Exposure Time (seconds)
ATG7 Blue ATG7 Rabbit Anti-Human
(78 kDa)
StarBright™ Blue 700 Goat Anti-Rabbit 6.9
AKR1C2 Red AKR1C2 Mouse Anti-Human (36 kDa) Goat Anti-Mouse IgG DyLight 800 300.0
Tubulin Green hFAB™ Rhodamine Anti-Tubulin
(50 kDa)
141.9


Detection of Post-Translational Modifications
Composite blot image
Composite blot image of triplex western blotting showing post-translational phosphorylation of a target protein.
Target Protein Color 1° Antibody 2° Antibody
Phospho-ERK 1/2
Red StarBright™ Blue 700 Goat Anti-Rabbit
Total ERK 1/2
Green Goat Anti-Mouse IgG DyLight 800
GAPDH Blue hFAB™ Rhodamine Anti-GAPDH


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StarBright Blue 700 Secondary Antibodies are ideal for fluorescent western blotting — either for the detection of a single target protein or for multiplex detection of several proteins on one blot, without stripping and reprobing. The StarBright Fluorophore is composed of a condensed polymer made up of multiple light-absorbing and -emitting monomers which provides an exceptionally bright signal (high quantum yield) compared to most traditional fluorophores.

StarBright Blue 700 Fluorescent Secondary Antibodies can be used with traditional fluorophores like the DyLight 800 Fluorophore for multiplexing. In addition, StarBright Antibodies can be used with Bio-Rad's stain-free technology and/or the hFAB™ Rhodamine Anti-Housekeeping Primary Antibodies for protein normalization. These antibodies are optimized for use with the ChemiDoc™ MP Imaging System, permitting detection of up to three proteins in a single blot.

Schematic representation of a StarBright Blue 700 Label conjugated to an antibody. The light-absorbing monomer is excited at 470 nm and, via an efficient intramolecular transfer process, the energy is transmitted to a light-emitting monomer, which emits at 700 nm. An antibody is conjugated to linker monomers.

Features and Benefits

  • Easy multiplexing — StarBright Secondary Antibodies can be used with IRDye 800 or other common fluorophores to detect multiple proteins on the same blot
  • Short exposure time — the exceptional brightness of the StarBright Fluorophore leads to shorter exposure times compared to traditional fluorescent dyes: seconds vs. minutes
  • High signal-to-noise ratio — StarBright Secondary Antibodies emit in the red/far red, where there is minimal background (Exmax/Emmax = 470 nm/700 nm)
  • Low nonspecific binding — StarBright Fluorophore is conjugated to highly cross-adsorbed antibodies

Applications and Uses

StarBright Blue 700 Secondary Antibodies can be used for multiplex fluorescent western blotting with the right combination of fluorophores as described in the table below. These fluorescent secondary antibodies can also be used to detect post-translational modifications like phosphorylation of a target protein.

Note: StarBright Antibodies are not suitable for stripping and reprobing.

Recommended Fluorophore Combinations
Targets Primary Target
(Fluorophore A)
Second Target
(Fluorophore B)
Third Target
(Fluorophore C)
1 StarBright Blue 700
2 StarBright Blue 700 hFAB Rhodamine HKP* Antibody (α-actin/tubulin/GAPDH)
3 StarBright Blue 700 DyLight 800 hFAB Rhodamine HKP Antibody (α-actin/tubulin/GAPDH)

* Stain-free gels can be used for total protein normalization in place of or in addition to housekeeping proteins (HKP).

Related Products

Related Categories

Supporting Documents

  • hFAB Rhodamine Housekeeping Antibodies Instruction Manual
  • Fluorescent Western Blotting with StarBright Blue 700 Secondary Antibodies Selection Guide
  • StarBright Blue 700 Secondary Antibodies Flier

Online Resources

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Waste Electrical and Electronic Equipment ("WEEE Directive")

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Waste Electrical and Electronic Equipment ("WEEE Directive")

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program

Waste Electrical and Electronic Equipment ("WEEE Directive")

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program

Waste Electrical and Electronic Equipment ("WEEE Directive")

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program

Waste Electrical and Electronic Equipment ("WEEE Directive")

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program

Goat Anti-Mouse IgG StarBright™ Blue 700, 80 µl
StarBright™ Blue 700 Goat Anti-Mouse IgG, 80 µl

12004159
Pkg of 1, lyophilized goat anti-mouse IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 80 µl

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$50.00
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StarBright™ Blue 700 Goat Anti-Mouse IgG, 400 µl
StarBright™ Blue 700 Goat Anti-Mouse IgG, 400 µl

12004158
Pkg of 1, lyophilized goat anti-mouse IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 400 µl

List Price:
 
$100.00
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StarBright™ Blue 700 Goat Anti-Mouse IgG, 5 x 400 µl
StarBright™ Blue 700 Goat Anti-Mouse IgG, 5 x 400 µl

12004157
Pkg of 5, lyophilized goat anti-mouse IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 400 µl

List Price:
 
$350.00
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StarBright™ Blue 700 Goat Anti-Rabbit IgG, 80 µl
StarBright™ Blue 700 Goat Anti-Rabbit IgG, 80 µl

12004162
Pkg of 1, lyophilized goat anti-rabbit IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 80 µl

List Price:
 
$50.00
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StarBright™ Blue 700 Goat Anti-Rabbit IgG, 400 µl
StarBright™ Blue 700 Goat Anti-Rabbit IgG, 400 µl

12004161
Pkg of 1, lyophilized goat anti-rabbit IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 400 µl

List Price:
 
$100.00
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StarBright™ Blue 700 Goat Anti-Rabbit IgG, 5 x 400 µl
StarBright™ Blue 700 Goat Anti-Rabbit IgG, 5 x 400 µl

12004160
Pkg of 5, lyophilized goat anti-rabbit IgG conjugated with StarBright Blue 700 Fluorophore; reconstitute to 400 µl

List Price:
 
$350.00
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Number Description Options
10000066837 StarBright™ Blue 700 Fluorescent Secondary Antibodies Instruction Manual, Ver B Click to download
10000078050 Instruction Manual, StarBright™ Blue 700 Secondary Antibodies and hFAB™ Rhodamine Anti‑Housekeeping Protein Primary Antibodies Click to download
10000068973 Fluorescent Western Blotting with StarBright™ Blue 700 Secondary Antibodies Selection Guide, Ver A Click to download
6911 StarBright™ Blue 700 Secondary Antibodies Flier Click to download