Nano-Secondary alpaca anti-rabbit IgG

Description
Nano-Secondary® Alpaca anti-rabbit IgG, recombinant VHH is an anti-rabbit IgG specific secondary antibody. It consists of of a mixture of 2 anti-rabbit Nanobodies that bind to rabbit IgG with high affinity & specificity. The product belongs to the Nano-Secondaries® family.

Target: Rabbit
Host: Alpaca-derived, recombinantly produced in bacteria
Specificity: Fab and Fc fragments of rabbit IgG
No cross-reactivity to: Mouse, rat, sheep, goat, and guinea pig serum
Cross-reactivity to: Human and macaque serum
Formats:
Conjugates: Alexa Fluor® 488, 568, and 647
Unconjugated: 2x Cystein

Applications
Immunofluorescence
Super-resolution microscopy
Western blotting
Flow cytometry

Fast one-step immunostaining
Simultaneous incubation of primary antibody and alpaca anti-rabbit IgG, recombinant VHH

Product Size Code Price Buy
Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] Size 10 µL Code srbAF488-1-10 Price $ 100
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Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102] Size 100 µL Code srbAF488-1-100 Price $ 365
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Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 568 [CTK0101, CTK0102] Size 10 µL Code srbAF568-1-10 Price $ 100
Buy +
Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 568 [CTK0101, CTK0102] Size 100 µL Code srbAF568-1-100 Price $ 365
Buy +
Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647 [CTK0101, CTK0102] Size 10 µL Code srbAF647-1-10 Price $ 100
Buy +
Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647 [CTK0101, CTK0102] Size 100 µL Code srbAF647-1-100 Price $ 365
Buy +
Product Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, for 2x Cys conjugation [CTK0101] Size 500 μg Code srbGCys2-1 Price Please inquire
Product Nano-Secondary® alpaca anti-rabbit IgG, recombinant VHH, for 2x Cys conjugation [CTK0102] Size 500 μg Code srbGCys2-2 Price Please inquire

Product type
Secondary Nanobody respectively VHH

Target/ Specificity
Fc [CTK0101] and Fab [CTK0102] fragments of rabbit IgG

Format
Alpaca single domain antibodies, monovalent

Host
Alpaca-derived, recombinantly produced in bacteria

Cross-reactivity
No cross-reactivity to mouse, rat, sheep, goat, and guinea pig IgG
Cross-reactivity to human and macaque IgG

Immunogen
Purified rabbit IgG

Clonality
Conjugated: Mixture of 2 monoclonal Nanobodies [CTK0101], [CTK0102]
Unconjugated: Monoclonal Nanobodies [CTK0101] or [CTK0102]

Clones
CTK0101, CTK0102

Conjugates
Site-directed conjugation to Alexa Fluor 488, Alexa Fluor 568, Alexa Fluor 647 or unconjugated

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.

Synonyms
Alpaca single domain antibody, VHH, Nanobody, binding domain of single domain antibody, Nano antibody

Validation
Application validated for immunofluorescence, SRM, and Western blotting
Determination of cross-reactivity, sequence, affinity, melting point, and degree of labeling (DOL)

Affinity (KD)
CTK0101: KD = 0.2 nM, CTK0102: KD = 1.2 nM

Degree of labeling (DOL)
2 fluorophores per Nanobody

Purity
Recombinantly expressed and purified

Form
Buffered aqueous solution

Storage buffer
10 mM HEPES pH 7.0, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % Sodium azide

Concentration
0.5 g/ L

Size
10 µL; 100 µL

How to cite these products
RRIDs:
Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488 [CTK0101, CTK0102]: AB_2827585
Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 568 [CTK0101, CTK0102]: AB_2827586
Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647 [CTK0101, CTK0102]: AB_2827587
Nano-Secondary® alpaca anti-human IgG/anti-rabbit IgG, recombinant VHH, for 2x Cys conjugation [CTK0101]: AB_2864261
Nano-Secondary® alpaca anti-rabbit IgG, recombinant VHH, for 2x Cys conjugation [CTK0102]: AB_2864262

Storage instructions
Shipped at ambient temperature. Store at -20°C short term or -80°C long term. Aliquot upon delivery. Avoid freeze-thaw cycles. Stable for 6 months after arrival.

Benefits

  • Reduced incubation and hands-on time because the Alpaca anti-rabbit IgG, recombinant VHH can be incubated together with the rabbit primary antibody
  • Higher resolution imaging due to the Alpaca anti-rabbit IgG, recombinant VHHs’ 10 times smaller size than conventional secondary antibodies. The small size enables better tissue penetration, antigen access and decreases the distance between the epitope and label.
  • Cleaner Images: The Alpaca anti-rabbit IgG, recombinant VHH has a high specificity and low background:
    We select only VHHs with the desired specificity during development; VHHs that cross-react to other commonly used species’ IgGs are eliminated. Hence, ChromoTek's Nano-Secondaries have an extreme low background; any kind of pre-adsorption is not needed.
  • Multiplexing: the Alpaca anti-rabbit IgG, recombinant VHH does not cross-react to other frequently used IgGs and makes multiplexing possible.

One-step immunostaining protocol saves time

ChromoTek’s Alpaca anti-rabbit IgG, recombinant VHH consists of monovalent Nanobodies with very high affinity. The monovalent Alpaca anti-rabbit IgG, recombinant VHH enables the simultaneous incubation together with rabbit primary antibodies.
This one-step immunostaining saves washing steps and incubation time. Simultaneous incubation also supports: 
•    multiplexing, 
•    tissue penetration, and
•    cell staining for flow cytometry.
Of course, Nano-Secondaries can be used according to standard immunostaining protocols following a sequential incubation of primary and Alpaca anti-rabbit IgG, recombinant VHH, too.

Better imaging

  • Better tissue penetration and small epitope-label displacement for higher resolution
  • High sensitivity: detection of low expressed epitopes via primary antibody
  • High affinity reduces background

No cross-reactivity to IgGs of other frequently used species

The cross-reactivity of ChromoTek’s Alpaca anti-rabbit IgG Nano-Secondary has been carefully tested. It is co-reactive to human IgG. It shows no cross-reactivity to mouse, rat, sheep, goat, and guinea pig serum. Cross-reactivity to macaque serum has been determined. Because of this high specificity our Nano-Secondaries do not require any kind of pre-adsorption.

Precise and reproducible staining

  • Site specific binding of 4 Alpaca anti-rabbit IgG, recombinant VHHs per rabbit primary IgG antibody
  • Stoichiometric labelling of 2 Alexa Fluor dyes per VHH respective Nanobody
  • Fully characterized and validated
  • Recombinantly produced, virtually without lot-to-lot variation
  • Manufacturing process is totally animal free, though VHHs respective Nanobodies are generated from immunization of alpacas and llamas.

Site-directed labeling of Nano-Secondaries

ChromoTek's Nano-Secondaries are conjugated to AlexaFluor 488, 568, or 647. For custom dyes, we offer unconjugated Nano-Secondaries. They have 2 Cystein residues for  conjugation with thiol-reactive dyes, e.g. maleimide. Please note that ChromoTek's Nano-Secondaries are often a mix of two different monoclonal Nanobodies whereas the unconjugated Nano-Secondaries are always sold as monoclonals and not pre-mixed.

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Only for research applications, not for diagnostic or therapeutic use!