Alpaca anti-rabbit IgG, recombinant VHH

Description
Nano-Secondaries are secondary antibodies. They are based on recombinant Nanobodies. Nano-Secondaries bind to rabbit IgG antibodies with high affinity & specificity.

Target: Rabbit
Host: Alpaca-derived, recombinantly produced in bacteria
Specificity: Fab and Fc fragments of rabbit IgG
Cross-reactivity to: Human and macaque
No cross-reactivity to: mouse, rat, sheep, goat, and guinea pig
Clonality: Biclonal, mixture of 2 monoclonal Nanobodies
Conjugates: Alexa Fluor® 488, 568, and 647

Applications
Immunofluorescence
Super-resolution microscopy
Western blot

Fast one-step immunostaining
Simultaneous incubation of primary antibody and rabbit Nano-Secondary

ProductSizeCodePriceBuy
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488Size10 µLCodesrbAF488-1-10Price $ 100
Buy+
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 488Size100 µLCodesrbAF488-1-100Price $ 365
Buy+
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 568Size10 µLCodesrbAF568-1-10Price $ 100
Buy+
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 568Size100 µLCodesrbAF568-1-100Price $ 365
Buy+
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647Size10 µLCodesrbAF647-1-10Price $ 100
Buy+
ProductAlpaca anti-rabbit IgG, recombinant VHH, Alexa Fluor® 647Size100 µLCodesrbAF647-1-100Price $ 365
Buy+

Product type
Secondary Nanobody

Target/ Specificity
Rabbit; this Nanobody mixture recognizes Fab and Fc fragments of rabbit IgG

Format
Alpaca single domain antibodies, monovalent

Host
Alpaca-derived, recombinantly produced in bacteria

Isotype
VHH domain of alpaca heavy chain antibody IgG2/3

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

Immunogen
Purified rabbit IgG

Clonality
Biclonal: mixture of 2 monoclonal Nanobodies

Clones
VHH0244, VHH0245

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

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)
VHH0244: KD = 0.2 nM, VHH0245: KD = 1.2 nM

Degree of labeling (DOL)
2 fluorophores per Nanobody

Purity
Recombinantly expressed and purified

Form
Buffered 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
Because of the small size of a VHH these volumes contain about 10 times more molecules than a comparable volume of conventional secondaries.

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

  • Higher resolution imaging
  • Reproducible results
  • Reduced incubation and hands-on time
  • Economic: only small amounts needed

One-step immunostaining protocol

Nano-Secondaries are monovalent antibodies that don’t cluster. Hence, they enable one-step immunostaining: the simultaneous incubation of primary and secondary antibodies. This optional method saves incubation time and washing steps.

Simultaneous incubation also supports multiplexing, i.e. the pre-incubation of multiple primary antibodies with Nano-secondaries conjugated to different fluorophores, which are then added to the sample. Even primaries from same subclass may be used.

Of course, Nano-Secondaries can be used according to standard immunostaining protocols following a sequential incubation of primary and secondary antibody, too.

Precise and reproducible staining

  • Recombinantly produced virtually without lot-to-lot variation
  • Stochiometric labelling of 2 Alexa Fluor dyes per Nanobody
  • Site specific binding of 4 Nano-Secondaries per primary rabbit IgG
  • Fully characterized and validated

Better imaging

  • Small epitope-label displacement for higher resolution
  • High sensitivity: detection of low expressed epitopes via primary antibody
  • High affinity reduces background

Cross reactivity

The cross reactivity of ChromoTek’s alpaca anti-rabbit Nano-Secondaries has been thoroughly tested against common secondary species: They show no cross-reactivity to mouse, rat, sheep, goat and guinea pig IgG & serum; cross-reactivity to human and macaque IgG & serum has been determined. For comparison, pre-adsorbed polyclonal secondary antibodies may show some levels of cross-reactivity.

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