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.

Reactivity

Rabbit, Human, Macaque

Applications

IF, WB, FC

Conjugate

Alexa Fluor® 488

Publications

4

Cat no : srbAF488-1

Synonyms

human IgG, rabbit IgG, immunoglobulin Gamma

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Validation Data Gallery

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  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.

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  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.

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  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.

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  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.

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  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.

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  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.

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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.

• Applications: IF, WB, FC
• Recommended Dilution: Immunofluorescence 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
• Affinity: CTK0101: KD = 0.2 nM, CTK0102: KD = 1.2 nM
• Specificity/Target: Human IgG and rabbit IgG
• Reactivity: Rabbit, Human, Macaque
• Conjugate: 0.5 g/ L
• Storage Buffer: 10 mM HEPES pH 7.0, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % Sodium azide
• Storage Condition: Store at -20°C short term or -80°C long term. Aliquot upon delivery. Avoid freeze-thaw cycles.

Publications

Application	Title
WB	Cancer Cell Int; CircFAM13B promotes the proliferation of hepatocellular carcinoma by sponging miR-212, upregulating E2F5 expression and activating the P53 pathway.; Authors - Ying Xie
	Anal Chem; Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination.; Authors - Mao Fukuyama
	Basic Res Cardiol; Inhibition of macrophage proliferation dominates plaque regression in response to cholesterol lowering.; Authors - Carmen Härdtner
IF	Nat Commun; Engineered macrophages as near-infrared light activated drug vectors for chemo-photodynamic therapy of primary and bone metastatic breast cancer.; Authors - Yanjuan Huang