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

Validation Data Gallery

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  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.

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  Dissociation constant Kd and affinity.

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  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.

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  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.

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  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.

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  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.

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  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.

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  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.

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  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.

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  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.

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.

Cat No. sms2bAF647-1

Publications(4)

Applications

IF, WB

Conjugate

Alexa Fluor® 647

Host

Alpaca

Reactivity

Mouse IgG2b

Synonyms

mouse IgG2b; mouse immunoglobulin Gamma 2b

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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.

• Description: Nano-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.
• Applications: IF, WB
• Host: Alpaca
• Species Reactivity: Mouse IgG2b Fc-fragment; No cross-reactivity: goat, guinea pig, rabbit, rat, sheep, human, macaque serum proteins, mouse IgG1, IgG2a, IgG2c, IgG3
• Target/Specificity: This Nanobody mixture recognizes the Fc fragment of mouse IgG2b.
• Physical State: Liquid
• Conjugate: Alexa Fluor® 647
• Suggested Dilution: Immunofluorescence 1:1,000; Western blot 1:1,000
• Purification Method: Recombinant expression, affinity purification IMAC
• Type: Mixture of 2 monoclonal Nanobodies
• Class: Recombinant
• Format: Alpaca single domain antibodies, monovalent
• Cross-reactivity: No cross-reactivity to rabbit, rat, sheep, goat, guinea pig, human, and macaque serum
• Immunogen: Purified mouse IgG2b
• Clonality: Biclonal: mixture of 2 monoclonal Nanobodies
• Clones: VHH0275, VHH0288
• RRID: AB_2827583
• Affinity (KD): CTK0105: K_D = 5 nM, CTK0106: K_D = 0.2 nM
• Excitation / Emission: Excitation max: 650 nm, Emission max: 665 nm
• Storage Buffer: 10 mM HEPES pH 7.0, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % Sodium azide
• Storage Condition: Store at -20°C short term or -80°C long term. Aliquot upon delivery. Avoid freeze-thaw cycles.
• Size: 10 µL; 100 µL
• Note: This product is for research use only, not for diagnostic or therapeutic use

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Publications

Application	Title
	Cell Mol Life Sci; SHANK3 deficiency leads to myelin defects in the central and peripheral nervous system.; Authors - Mariagiovanna Malara
IF	Eur J Pharmacol; P2Y14 receptor in trigeminal ganglion contributes to neuropathic pain in mice; Authors - Jiu Lin
IF	Breast Cancer Res; UCHL1 contributes to insensitivity to endocrine therapy in triple-negative breast cancer by deubiquitinating and stabilizing KLF5; Authors - Juan Li
IF	ACS Omega; The Development and Characterization of Two Monoclonal Antibodies Against the Conjugates and Derivatives of the Immunometabolite Itaconate; Authors - Lauren Switala