Alpaca anti-mouse IgG2b, recombinant VHH

Description
Alpaca anti-mouse IgG2b, Fc-specific recombinant VHH is an anti-mouse IgG subclass specific antibody. This secondary antibody product consists of Nanobodies that bind to mouse IgG2b with high affinity & specificity.

Target: Mouse IgG2b
Host: Alpaca-derived, recombinantly produced in bacteria
Specificity: Fc fragments of mouse IgG2b
No cross-reactivity to: Rabbit, rat, goat, sheep, guinea pig, human, and macaque serum and to mouse IgG1, IgG2a, IgG2c, and IgG3
Clonality: Biclonal, mixture of 2 monoclonal Nanobodies
Conjugates: Alexa Fluor® 647

Applications
Immunofluorescence
Super-resolution microscopy
Western blotting

Fast one-step immunostaining
Simultaneous incubation of mouse IgG2b primary antibody and Alpaca anti-mouse IgG2b, recombinant VHH

Product Size Code Price Buy
Product Alpaca anti-mouse IgG2b, Fc-specific recombinant VHH, Alexa Fluor® 647 Size 10 µL Code sms2bAF647-1-10 Price $ 100
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Product Alpaca anti-mouse IgG2b, Fc-specific recombinant VHH, Alexa Fluor® 647 Size 100 µL Code sms2bAF647-1-100 Price $ 365
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Product type
Secondary Nanobody (secondary VHH)

Target/ Specificity
Mouse IgG2b; this Nanobody mixture recognizes the Fc fragment of mouse IgG2b

Format
Alpaca single domain antibodies, monovalent

Host
Alpaca-derived, recombinantly produced

Cross-reactivity
No cross-reactivity to: rabbit, rat, goat, sheep, guinea pig, human, and macaque serum and to mouse IgG1, IgG2a, IgG2c, and IgG3

Immunogen
Purified mouse IgG2b

Clonality
Biclonal: mixture of 2 monoclonal Nanobodies

Clones
VHH0275, VHH0288

Conjugates
Site-directed conjugation to Alexa Fluor 647

Recommended dilution
Immunofluorescence 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, subclass specificity, sequence, affinity, melting point, and degree of labeling (DOL)

Affinity (KD)
VHH0275: KD = 5 nM, VHH0288: KD = 0.2 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

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

Faster staining
One-step staining saves time due to simultaneous incubation of mouse IgG2b primary antibody and Alpaca anti-mouse IgG2b, recombinant VHH.

Higher resolution
Alpaca anti-mouse IgG2b, recombinant VHH is about 10 times smaller than conventional secondary antibodies. The small size enables better tissue penetration, antigen access, and has a small epitope to label displacement.

Cleaner Images
Nano-Secondaries have a high specificity and low background:

We select only VHHs with the desired specificity during development; VHHs that cross-react to IgGs from other commonly used species and to other IgG subclasses are eliminated. Therefore, our Nano-Secondaries have a very low background and hence do not require any kind of pre-adsorption.

Multiplexing
Alpaca anti-mouse IgG2b, recombinant VHH is subclass-specific and does not cross-react to IgGs from other commonly used species or other mouse IgG subclasses. This high specificity in combination with low background enables multiplexing.

Time savings by one-step immunostaining

ChromoTek’s Alpaca anti-mouse IgG2b, recombinant VHH consists of 2 monovalent Nanobodies with very high affinity. Hence, they enable one-step immunostaining: the simultaneous incubation of mouse IgG2b primary antibody and Alpaca anti-mouse IgG2b, recombinant VHH.

This method saves incubation time and washing steps.

Of course, Nano-Secondaries can be used according to standard immunostaining protocols following a sequential incubation of mouse IgG2b primary antibody and Alpaca anti-mouse IgG2b, recombinant VHH, too.

Simultaneous incubation also supports:

  • multiplexing
  • tissue penetration
  • cell staining for flow cytometry

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 IgG from other commonly used species and mouse IgG subclasses

The cross-reactivity of ChromoTek’s Alpaca anti-mouse IgG2b Nano-Secondary has been carefully tested. It shows no cross-reactivity to the mouse subclasses IgG1, IgG2a, IgG2c, and IgG3, and no cross-reactivity to rabbit, rat, goat, sheep, guinea pig, human, and macaque serum.

Precise and reproducible staining

  • Site specific binding of 4 Nanobodies per mouse IgG2b primary antibody
  • Stoichiometric labelling of 2 Alexa Fluor dyes per Nanobody
  • Characterized and validated: DNA and protein sequences, affinity, melting temperature
  • Recombinantly produced, virtually without lot-to-lot variation
  • Manufacturing process is totally animal free, though Nanobodies are generated from immunization of alpacas and llamas.

 

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