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ChromoTek Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102]

Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH is an anti-human IgG and anti-rabbit IgG specific secondary antibody. It consists of of a mixture of 2 Nanobodies that bind to human IgG and rabbit IgG with high affinity & specificity.

Ready to use reagent

Reactivity

Rabbit, Human, Macaque

Applications

IF, WB, FC

Conjugate

Alexa Fluor® 488

Cat no : srbAF488-1

Synonyms

human IgG, rabbit IgG, immunoglobulin Gamma

Validation Data Gallery

  • Alpaca anti-rabbit IgG VHH Alexa Fluor® 488 was applied in conjunction with rabbit anti-Lamin B1 antibodies for immunostaining of nuclear lamina (green) in HeLa cells. Scale bar, 20 µm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Alpaca anti-rabbit IgG VHH Alexa Fluor® 488 was applied in conjunction with rabbit anti-Lamin B1 antibodies for immunostaining of nuclear lamina (green) in HeLa cells. Scale bar, 20 µm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Higher resolution with anti-rabbit IgG Nano-Secondaries compared to conventional secondary antibodies: Left: Formation of a small, precise complex of Nanobodies (green) & primary antibody (grey). Right: Formation of a large, arbitrary complex of multiple polyclonal secondaries (green) & primary rabbit antibody.

    Higher resolution with anti-rabbit IgG Nano-Secondaries compared to conventional secondary antibodies: Left: Formation of a small, precise complex of Nanobodies (green) & primary antibody (grey). Right: Formation of a large, arbitrary complex of multiple polyclonal secondaries (green) & primary rabbit antibody.

  • Well-defined and characterized immunostaining: Primary anti-rabbit IgG antibody (grey) with 2 copies each of a rabbit Fab- and Fc-specific monoclonal Nanobodies (green) bound. In total, 8 fluorophores (red stars) label the primary rabbit IgG antibody.

    Well-defined and characterized immunostaining: Primary anti-rabbit IgG antibody (grey) with 2 copies each of a rabbit Fab- and Fc-specific monoclonal Nanobodies (green) bound. In total, 8 fluorophores (red stars) label the primary rabbit IgG antibody.

  • HeLa cells were immunostained with rabbit anti-Lamin B1 antibodies and alpaca anti-rabbit IgG VHH Alexa Fluor® 568 (1:1,000). Confocal and gated STED images were acquired with a Leica TCS SP8 STED 3X microscope, pulsed depletion with a 775 nm laser. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    HeLa cells were immunostained with rabbit anti-Lamin B1 antibodies and alpaca anti-rabbit IgG VHH Alexa Fluor® 568 (1:1,000). Confocal and gated STED images were acquired with a Leica TCS SP8 STED 3X microscope, pulsed depletion with a 775 nm laser. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • Multiple Nano-Secondaries can be applied for multiplex fluorescent Western blotting. This allows multiple targets to be analyzed simultaneously on the same blot at the same time. Multiplex fluorescent Western blot of GFP-TOM70, ®-Tubulin, and GFP in HEK293T cell lysate. Western blot membrane was simultaneously incubated with primary antibodies and Nano-Secondaries. Green: rabbit anti-GFP (ChromoTek PABG1) + alpaca anti-rabbit IgG VHH Alexa Fluor® 488. Magenta: mouse anti-®-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

    Multiple Nano-Secondaries can be applied for multiplex fluorescent Western blotting. This allows multiple targets to be analyzed simultaneously on the same blot at the same time. Multiplex fluorescent Western blot of GFP-TOM70, ®-Tubulin, and GFP in HEK293T cell lysate. Western blot membrane was simultaneously incubated with primary antibodies and Nano-Secondaries. Green: rabbit anti-GFP (ChromoTek PABG1) + alpaca anti-rabbit IgG VHH Alexa Fluor® 488. Magenta: mouse anti-®-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

  • Western blot analysis of EGFP (EGFP-250, ChromoTek) added to HEK293T cell lysate. Detection with rabbit anti-GFP antibody PABG1 (ChromoTek) and alpaca anti-rabbit IgG VHH Alexa Fluor® 488.

    Western blot analysis of EGFP (EGFP-250, ChromoTek) added to HEK293T cell lysate. Detection with rabbit anti-GFP antibody PABG1 (ChromoTek) and alpaca anti-rabbit IgG VHH Alexa Fluor® 488.

Product Information

Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH is an anti-human IgG and anti-rabbit IgG specific secondary antibody. It consists of of a mixture of 2 Nanobodies that bind to human IgG and rabbit IgG with high affinity & specificity.

ApplicationsIF, WB, FC
Recommended DilutionImmunofluorescence 1:1,000
Super-resolution microscopy 1:1,000
Western blot 1:1,000
Optimal working concentration is application-dependent and should be determined by testing a dilution range from 1:250 to 1:2,000.
Note: Image acquisition time may have to be optimized
Clonality Biclonal: mixture of 2 monoclonal Nanobodies
Host Alpaca
AffinityCTK0101: KD = 0.2 nM, CTK0102: KD = 1.2 nM
Specificity/TargetHuman IgG and rabbit IgG
ReactivityRabbit, Human, Macaque
ConjugateAlexa Fluor® 488
Storage Buffer10 mM HEPES pH 7.0, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % Sodium azide
Storage ConditionStore at -20°C short term or -80°C long term. Aliquot upon delivery. Avoid freeze-thaw cycles.

Publications

ApplicationTitle

Anal Chem

Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination.

Authors - Mao Fukuyama
View Article

Basic Res Cardiol

Inhibition of macrophage proliferation dominates plaque regression in response to cholesterol lowering.

Authors - Carmen Härdtner
View Article