Alpaca anti-mouse IgG1, recombinant VHH
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Grey: Mouse IgG1 anti-Vimentin + alpaca anti-mouse IgG1VHH Alexa Fluor 647. Green: Mouse IgG2b anti-Lamin + alpaca anti-mouse IgG2b VHH Alexa Fluor 488.
Red: Mouse IgG3 anti-MOT + alpaca anti-mouse IgG3 VHH Alexa Fluor 568.
Scale bar, 10 µm.
Scale bar, 20 µm.
Grey: Mouse IgG1 anti-Vimentin + alpaca anti-mouse IgG1VHH Alexa Fluor 647. Green: Mouse IgG2b anti-Lamin + alpaca anti-mouse IgG2b VHH Alexa Fluor 488.
Red: Mouse IgG3 anti-MOT + alpaca anti-mouse IgG3 VHH Alexa Fluor 568.
Scale bar, 10 µm.
Description
Alpaca anti-mouse IgG1, Fc-specific recombinant VHH is an anti-mouse IgG subclass specific antibody. This secondary antibody product consists of Nanobodies that bind to mouse IgG1 with high affinity & specificity.
Target: Mouse IgG1
Host: Alpaca-derived, recombinantly produced in bacteria
Specificity: Fc fragment of mouse IgG1
No cross-reactivity to: Goat, guinea pig, rabbit, rat, sheep, human, and macaque serum and to mouse IgG2a, IgG2b, 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 IgG1 primary antibody and Alpaca anti-mouse IgG1, recombinant VHH
| Product | Size | Code | Price | Buy |
|---|---|---|---|---|
| Product Alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 647 | Size 10 µL | Code sms1AF647-1-10 | Price $ 100 |
Buy
+
|
| Product Alpaca anti-mouse IgG1, recombinant VHH, Alexa Fluor® 647 | Size 100 µL | Code sms1AF647-1-100 | Price $ 365 |
Buy
+
|
Product type
Secondary Nanobody (secondary VHH)
Target/ Specificity
Mouse IgG1; this Nanobody mixture recognizes Fc fragments of mouse IgG1
Format
Alpaca single domain antibodies, monovalent
Host
Alpaca-derived, recombinantly produced in bacteria
Cross-reactivity
No cross-reactivity to rabbit, rat, goat, sheep, guinea pig, human, and macaque serum and to mouse IgG2a, IgG2b, IgG2c, and IgG3
Immunogen
Purified mouse IgG1
Clonality
Biclonal: mixture of 2 monoclonal Nanobodies
Clones
VHH302, VHH305
Conjugates
Site-directed conjugation to Alexa Fluor 647
Recommended dilution
Immunofluorescence 1:500
Super-resolution microscopy 1:500
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)
VHH302: KD =0.13 nM, VHH305: KD = 0.63nM
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 IgG1 primary antibody and Alpaca anti-mouse IgG1, recombinant VHH.
Higher resolution
Alpaca anti-mouse IgG1, recombinant VHH is about 10 times smaller than conventional secondary antibodies. The small size enables better tissue penetration, antigen access, and decreases the distance between epitope and label.
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 IgG1, 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 IgG1, recombinant VHH consists of 2 monovalent Nanobodies with very high affinity. Hence, they enable one-step immunostaining: the mouse IgG1 primary antibody and alpaca anti-mouse IgG1, recombinant VHH can be incubated together.
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 IgG1 primary antibody and alpaca anti-mouse IgG1, 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-expression 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 IgG1 Nano-Secondary has been carefully tested. It shows no cross-reactivity to the mouse subclasses IgG2a, IgG2b, 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 IgG1 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.
Only for research applications, not for diagnostic or therapeutic use!