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ChromoTek Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, Alexa Fluor® 647 [CTK0105, CTK0106]

Nano-Secondary® 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.

Applications

IF, WB

Conjugate

Alexa Fluor® 647

Host

Alpaca

Reactivity

Mouse IgG2b

Cat no : sms2bAF647-1

Synonyms

mouse IgG2b; mouse immunoglobulin Gamma 2b

Validation Data Gallery

  • Anti-mouse IgG2b Nano-Secondary: Well-defined and characterized immunostaining. Primary anti-mouse IgG2b antibody (grey) with 2X2 monoclonal mouse Fc- specific Nanobodies (green) bound. In total, 8 fluorophores (red stars) label the mouse IgG2b primary antibody.

    Anti-mouse IgG2b Nano-Secondary: Well-defined and characterized immunostaining. Primary anti-mouse IgG2b antibody (grey) with 2X2 monoclonal mouse Fc- specific Nanobodies (green) bound. In total, 8 fluorophores (red stars) label the mouse IgG2b primary antibody.

  • Dissociation constant Kd and affinity.

    Dissociation constant Kd and affinity.

  • Multiplexed immunostaining of HeLa cells with two alpaca anti-mouse Nano-Secondaries and one anti-rabbit Nano-Secondary. Green: mouse IgG1 anti-COX4 + alpaca anti-mouse IgG1 VHH Alexa Fluor® 488. Magenta: mouse IgG2b anti-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647. Yellow: rabbit anti-Lamin + alpaca anti-rabbit IgG VHH Alexa Fluor® 568. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Multiplexed immunostaining of HeLa cells with two alpaca anti-mouse Nano-Secondaries and one anti-rabbit Nano-Secondary. Green: mouse IgG1 anti-COX4 + alpaca anti-mouse IgG1 VHH Alexa Fluor® 488. Magenta: mouse IgG2b anti-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647. Yellow: rabbit anti-Lamin + alpaca anti-rabbit IgG VHH Alexa Fluor® 568. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • The anti-mouse IgG2b Nano-Secondary is subclass-specific and does not cross-react with IgGs from other commonly used species (here rabbit) and with mouse IgG1 and IgG3 subclasses.

    The anti-mouse IgG2b Nano-Secondary is subclass-specific and does not cross-react with IgGs from other commonly used species (here rabbit) and with mouse IgG1 and IgG3 subclasses.

  • One-step staining (left) vs. sequential staining (right) of HeLa cells with anti-Lamin A/C (nuclear lamina) mouse IgG2b monoclonal primary antibody + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647 (magenta). Scale bar, 20 μm.

    One-step staining (left) vs. sequential staining (right) of HeLa cells with anti-Lamin A/C (nuclear lamina) mouse IgG2b monoclonal primary antibody + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647 (magenta). Scale bar, 20 μm.

  • Multiplexed immunostaining of HeLa cells with two alpaca anti-mouse Nano-Secondaries and one anti-rabbit Nano-Secondary. Green: mouse IgG1 anti-COX4 + alpaca anti-mouse IgG1 VHH Alexa Fluor® 488. Yellow: rabbit anti-Lamin + alpaca anti-rabbit IgG VHH Alexa Fluor® 568. Magenta: mouse IgG2b anti-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    Multiplexed immunostaining of HeLa cells with two alpaca anti-mouse Nano-Secondaries and one anti-rabbit Nano-Secondary. Green: mouse IgG1 anti-COX4 + alpaca anti-mouse IgG1 VHH Alexa Fluor® 488. Yellow: rabbit anti-Lamin + alpaca anti-rabbit IgG VHH Alexa Fluor® 568. Magenta: mouse IgG2b anti-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

  • One-step immunostaining is the simultaneous incubation of mouse IgG2b primary antibody and anti-mouse IgG2b Nano-Secondary. This method reduces incubation and hands-on time. Simultaneous incubation also supports multiplexing, tissue penetration, and cell staining for flow cytometry.

    One-step immunostaining is the simultaneous incubation of mouse IgG2b primary antibody and anti-mouse IgG2b Nano-Secondary. This method reduces incubation and hands-on time. Simultaneous incubation also supports multiplexing, tissue penetration, and cell staining for flow cytometry.

  • Multiple Nano-Secondaries can be applied for multiplex fluorescent Western blotting. This allows multiple targets to be analyzed simultaneously on the same blot at the same time. Multiplex fluorescent Western blot of GFP-TOM70, ®-Tubulin, and GFP in HEK293T cell lysate. Western blot membrane was simultaneously incubated with primary antibodies and Nano-Secondaries. Green: rabbit anti-GFP (ChromoTek PABG1) + alpaca anti-rabbit IgG VHH Alexa Fluor® 488. Magenta: mouse anti-®-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

    Multiple Nano-Secondaries can be applied for multiplex fluorescent Western blotting. This allows multiple targets to be analyzed simultaneously on the same blot at the same time. Multiplex fluorescent Western blot of GFP-TOM70, ®-Tubulin, and GFP in HEK293T cell lysate. Western blot membrane was simultaneously incubated with primary antibodies and Nano-Secondaries. Green: rabbit anti-GFP (ChromoTek PABG1) + alpaca anti-rabbit IgG VHH Alexa Fluor® 488. Magenta: mouse anti-®-Tubulin + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

  • Western blot analysis of endogenous ®-Tubulin in HEK293T cell lysate. Detection with mouse anti-®-Tubulin antibody and alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

    Western blot analysis of endogenous ®-Tubulin in HEK293T cell lysate. Detection with mouse anti-®-Tubulin antibody and alpaca anti-mouse IgG2b VHH Alexa Fluor® 647.

  • HeLa cells were immunostained with mouse IgG2b anti-®-Tubulin antibody + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647 (magenta). Nuclei were stained with DAPI, blue. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

    HeLa cells were immunostained with mouse IgG2b anti-®-Tubulin antibody + alpaca anti-mouse IgG2b VHH Alexa Fluor® 647 (magenta). Nuclei were stained with DAPI, blue. Scale bar, 10 μm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

Product Information

Nano-Secondary® 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.

DescriptionNano-Secondary® alpaca anti-mouse IgG2b, Fc-specific recombinant VHH reagent uses a novel class of anti-mouse IgG isotype specific antibodies This secondary antibody product consists of Nanobodies that bind to mouse IgG2b with high affinity & specificity. Nano-Secondary reagents enable simultaneous incubation with primary antibodies, thus shortening your experimental. Due to their small size, Nano-Secondary reagents provide enhanced image resolution. No cross-reactivity against serum proteins from other species, details see below.
ApplicationsIF, WB
HostAlpaca
Species ReactivityMouse IgG2b Fc-fragment
No cross-reactivity: goat, guinea pig, rabbit, rat, sheep, human, macaque serum proteins, mouse IgG1, IgG2a, IgG2c, IgG3
Physical StateLiquid
ConjugateAlexa Fluor® 647
Suggested DilutionImmunofluorescence 1:1,000
Western blot 1:1,000
Purification MethodRecombinant expression, affinity purification IMAC
TypeMixture of 2 monoclonal Nanobodies
ClassRecombinant
Affinity (KD) CTK0105: KD = 5 nM, CTK0106: KD = 0.2 nM
Storage Buffer10 mM HEPES pH 7.0, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % Sodium azide
Storage ConditionStore at -20°C short term or -80°C long term. Aliquot upon delivery. Avoid freeze-thaw cycles.
RRIDAB_2827583
NoteThis product is for research use only, not for diagnostic or therapeutic use

Publications

ApplicationTitle

Cell Mol Life Sci

SHANK3 deficiency leads to myelin defects in the central and peripheral nervous system.

Authors - Mariagiovanna Malara
View Article
IF

Eur J Pharmacol

P2Y14 receptor in trigeminal ganglion contributes to neuropathic pain in mice

Authors - Jiu Lin
View Article