ChromoTek Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647 [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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  HeLa cells stably expressing Tubulin-GFP at near-endogenous level were immunostained with rabbit anti-GFP PABG1 antibody and alpaca anti-rabbit IgG VHH Alexa Fluor® 647 (magenta). Nuclei were detected with H2B-RFP and RFP-Booster Atto594 (cyan). Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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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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  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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  One-step immunostaining (top row) vs. sequential immunostaining (bottom row) of HeLa cells. Anti-β-Actin (left column) and anti-LaminB1 (right column) primary rabbit antibodies + secondary alpaca anti-rabbit IgG VHH Alexa Fluor® 647. Scale bar, 20 μm.

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. srbAF647-1

Publications(3)

Reviews (2)

Applications

IF, WB, FC

Reactivity

Rabbit, Human, Macaque

Host

Alpaca

Conjugate

Alexa Fluor® 647

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® 647
• 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_2827587
• Excitation / Emission: Excitation max: 650 nm, Emission max: 665 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	Biosens Bioelectron; 3D co-culture of macrophages and fibroblasts in a sessile drop array for unveiling the role of macrophages in skin wound-healing; Authors - Xiaoyan Lyu
IF	Vet Microbiol; NDV inhibited IFN-β secretion through impeding CHCHD10-mediated mitochondrial fusion to promote viral proliferation; Authors - Xibing Yu
IF	Nat Methods; Spatial single-cell mass spectrometry defines zonation of the hepatocyte proteome; Authors - Florian A Rosenberger

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Clarisse (Verified Customer) (08-08-2022)

This secondary nano-body performs incredibly well in both single-step and sequential IF incubations. The dilution used was 1:1000. The signal is way more specific compared to conventional alexa secondaries. You should definitely give it a try!

• Applications: Immunofluorescence
• Cell Tissue Type: Human cerebral organoids

FH

Joe (Verified Customer) (04-26-2022)

I used this secondary in an indirect flow cytometry experiment, staining for a phospho-protein. I found that this secondary resulted in a stronger signal when compared to my regular life scientific alexa fluor secondary antibody. I will be using these nanobodies from now on! The antibody was used at a dilution of 1:4000, 30m RT staining.

• Applications: Flow Cytometry
• Cell Tissue Type: K562