ChromoTek Nano-Secondary® anti-mouse IgG1, recombinant VHH, Alexa Fluor® 647 [CTK0103, CTK0104]

Validation Data Gallery

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  Anti-mouse IgG1 Nano-Secondary: Well-defined and characterized immunostaining. Primary anti-mouse IgG1 antibody (grey) with 2X2 monoclonal mouse Fc- specific Nanobodies (green) bound. In total, 8 fluorophores (red stars) label the mouse IgG1 primary antibody.

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  Immunostaining with Alpaca anti-mouse IgG1 Alexa Fluor 647 Nano-Secondaries for FACS. Jurkat CD3+ and Jurkat CD3- cell lines were mixed and immunostained live with anti-CD3 mouse IgG1 + alpaca anti-mouse IgG1 VHH Alexa Fluor 647 (1:600). CD3- cells were pre-stained with Calcein. Two cell populations can be clearly distinguished on the dot-plot: Alexa Fluor 647-positive Calcein-negative cells and Alexa Fluor 647-negative Calcein-positive cells.

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  Multiplexed immunostaining of HeLa cells with 3 subclass-specific alpaca anti-mouse Nano-Secondaries. Grey: Mouse IgG1 anti-Vimentin + alpaca anti-mouse IgG1 VHH 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. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  Western blot analysis of EGFP (EGFP-250, ChromoTek) added to HEK293T cell lysate. Detection with anti-GFP mouse IgG 1 antibody and alpaca anti-mouse IgG1 VHH Alexa Fluor® 647.

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  One-step staining (left) vs. sequential staining (right) of HeLa cells with anti-COX4 (mitochondria) mouse IgG1 monoclonal primary antibody + alpaca anti-mouse IgG1 VHH Alexa Fluor® 647 (magenta). Cell nuclei are stained with DAPI (blue). Scale bar, 20 μm.

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

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  Multiplexed immunostaining of HeLa cells with 3 subclass-specific alpaca anti-mouse Nano-Secondaries. Grey: Mouse IgG1 anti-Vimentin + alpaca anti-mouse IgG1 VHH 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. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

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  HeLa cells were immunostained with anti-Vimentin mouse IgG1 antibody and alpaca anti-mouse IgG1 VHH Alexa Fluor® 647 (grey). Nuclei were stained with DAPI, blue. 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 IgG1 primary antibody and anti-mouse IgG1 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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  Flow cytometry with anti-mouse Nano-Secondaries. Jurkat CD3+ and Jurkat CD3- cell lines were mixed and immunostained live with anti-CD3 mouse IgG1 and alpaca anti-mouse IgG1 VHH Alexa Fluor® 647 (1:600). Two cell populations can be clearly distinguished with a 2-log shift: CD3-positive cells (in green) and CD3-negative cells (in grey).

Nano-Secondary® anti-mouse IgG1, 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 IgG1 with high affinity & specificity.

Cat No. sms1AF647-1

Publications(3)

Applications

IF, WB, FC

Conjugate

Alexa Fluor® 647

Host

Alpaca

Reactivity

Mouse IgG1

Synonyms

mouse IgG1; mouse immunoglobulin Gamma 1

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Product Information

Nano-Secondary® anti-mouse IgG1, 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 IgG1 with high affinity & specificity.

• Description: Nano-Secondary® alpaca anti-mouse IgG1, 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 IgG1 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, FC
• Host: Alpaca
• Species Reactivity: Mouse IgG1 Fc-fragment; No cross-reactivity: goat, guinea pig, rabbit, rat, sheep, human, macaque serum proteins, mouse IgG2a, IgG2b, IgG2c, IgG3
• Target/Specificity: This Nanobody mixture recognizes the Fc fragment of mouse IgG1.
• Physical State: Liquid
• Conjugate: Alexa Fluor® 647
• Suggested Dilution: Immunofluorescence 1:500; Super-resolution microscopy 1:500; 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 IgG1
• Clonality: Biclonal: mixture of 2 monoclonal Nanobodies
• Clones: VHH0302, VHH0305
• RRID: AB_2827579
• Affinity (KD): CTK0103: K_D =0.13 nM, CTK0104: K_D = 0.63nM
• 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
IF	Gene Ther; A new protocol for whole-brain biodistribution analysis of AAVs by tissue clearing, light-sheet microscopy and semi-automated spatial quantification; Authors - Miguel M Lopes
	Cell Rep; Hypothalamic astrocytic-BMAL1 regulates energy homeostasis in a sex-dependent manner; Authors - María Luengo-Mateos
	PLoS Pathog; Peculiar transcriptional reprogramming with functional impairment of dendritic cells upon exposure to transformed HTLV-1-infected cells; Authors - Auriane Carcone