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

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.

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.

Cat No. srbAF488-1

Publications(47)

Applications

IF, WB, FC

Reactivity

Rabbit, Human, Macaque

Host

Alpaca

Conjugate

Alexa Fluor® 488

Synonyms

human IgG, rabbit IgG, immunoglobulin Gamma

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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.

• Description: Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH reagent uses a novel class of anti-rabbit IgG and anti-human IgG specific antibodies. This secondary antibody product consists of Nanobodies that bind to rabbit IgG and human IgG with high affinity & specificity. Nano-Secondary reagents enable simultaneous incubation with primary antibodies, thus shortening your experimental. Due to their small size, Nano-Secondary reagents provide enhanced image resolution. No cross-reactivity against serum proteins from other species, details see below.
• Applications: IF, WB, FC
• Host: Alpaca
• Species Reactivity: Rabbit, Human, Macaque; No cross-reactivity to mouse, rat, sheep, goat, and guinea pig IgG
• Target/Specificity: Fc-fragment of human IgG, Fab- and Fc-fragment of rabbit IgG (co-reactivity)
• Physical State: Liquid
• Conjugate: Alexa Fluor® 488
• Suggested Dilution: Immunofluorescence 1:1,000; Super-resolution microscopy 1:1,000; Western blot 1:1,000
• Purification Method: Recombinant expression, affinity purification IMAC
• Type: Mixture of 2 monoclonal Nanobodies; Secondary Nanobody
• Class: Recombinant
• Format: Alpaca single domain antibody, monovalent
• Cross-reactivity: No cross-reactivity to goat, guinea pig, mouse, rat, and sheep serum; Cross-reactivity to macaque serum
• Immunogen: Purified rabbit IgG
• Clonality: Biclonal: mixture of 2 monoclonal Nanobodies
• Clones: CTK0101 (VHH0244): Fc-fragment of human IgG, Fc-fragment of rabbit IgG; CTK0102 (VHH0245): Fab-fragment of rabbit IgG
• Affinity (KD): CTK0101: K_D = 0.2 nM, CTK0102: K_D = 1.2 nM
• RRID: AB_2827585
• Excitation / Emission: Excitation max: 490 nm, Emission max: 525 nm
• 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.
• Size: 10 μL; 100 μL
• Note: This product is for research use only, not for diagnostic or therapeutic use

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Publications

Application	Title
IF	Sci Adv; Mutation of SLC7A14 causes auditory neuropathy and retinitis pigmentosa mediated by lysosomal dysfunction.; Authors - Kimberlee P Giffen
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
	Basic Res Cardiol; Inhibition of macrophage proliferation dominates plaque regression in response to cholesterol lowering.; Authors - Carmen Härdtner
IF	Biomed Pharmacother; Rutaecarpine alleviates inflammation and fibrosis by targeting CK2α in diabetic nephropathy; Authors - Juan Chen
	Anal Chem; Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination.; Authors - Mao Fukuyama
IF	Front Pharmacol; Design and synthesis of cabotegravir derivatives bearing 1,2,3-triazole and evaluation of anti-liver cancer activity; Authors - Huaxia Xie