Nano-Secondary alpaca anti-mouse IgG2b

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. The product belongs to the Nano-Secondaries® family.

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
Formats:
Conjugated: Alexa Fluor® 488, 568 and 647
Unconjugated:  2x Cystein

Applications
Immunofluorescence
Super-resolution microscopy
Western blotting

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

Product Size Code Price Buy
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 488 [CTK0105, CTK0106] Size 10 µL Code sms2bAF488-1-10 Price $ 100
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 488 [CTK0105, CTK0106] Size 100 µL Code sms2bAF488-1-100 Price $ 365
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 568 [CTK0105, CTK0106] Size 10 µL Code sms2bAF568-1-10 Price $ 100
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 568 [CTK0105, CTK0106] Size 100 µL Code sms2bAF568-1-100 Price $ 365
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 647 [CTK0105, CTK0106] Size 10 µL Code sms2bAF647-1-10 Price $ 100
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 647 [CTK0105, CTK0106] Size 100 µL Code sms2bAF647-1-100 Price $ 365
Buy +
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, for 2x Cys conjugation [CTK0105] Size 500 μg Code smsG2bCys2-1 Price Please inquire
Product Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, for 2x Cys conjugation [CTK0106] Size 500 μg Code smsG2bCys2-2 Price Please inquire

Product type
Secondary Nanobody (secondary VHH)

Target/ Specificity
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
Conjugated: Mixture of 2 monoclonal Nanobodies [CTK0105], [CTK0106]
Unconjugated: Monoclonal Nanobodies [CTK0105] or [CTK0106]

Clones
CTK0105, CTK0106

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

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)
CTK0105: KD = 5 nM, CTK0106: KD = 0.2 nM

Degree of labeling (DOL)
2 fluorophores per Nanobody

Purity
Recombinantly expressed and purified

Form
Buffered aqueous solution

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

Concentration
0.5 g/ L

 

How to cite these products
RRIDs:
Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 488 [CTK0105, CTK0106]: AB_2827581
Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 568 [CTK0105, CTK0106]: AB_2827582
Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 647 [CTK0105, CTK0106]: AB_2827583
Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, for 2x Cys conjugation [CTK0105]: AB_2865265
Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, for 2x Cys conjugation [CTK0106]: AB_2864266

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.

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!