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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.

Ready to use reagent

Reactivity

Mouse

Applications

IF, WB

Conjugate

Alexa Fluor® 647

Cat no : sms2bAF647-1

Synonyms

mouse IgG2b; mouse immunoglobulin Gamma 2b

Validation Data Gallery

  • 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.

  • 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.

  • 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.

ApplicationsIF, WB
Recommended DilutionImmunofluorescence 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.
Clonality Biclonal: mixture of 2 monoclonal Nanobodies
Host Alpaca
AffinityCTK0105: KD = 5 nM, CTK0106: KD = 0.2 nM
Specificity/TargetMouse IgG2b; this Nanobody mixture recognizes the Fc fragment of mouse IgG2b
ReactivityMouse
ConjugateAlexa Fluor® 647
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.