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

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.

Applications

IF, WB, FC

Conjugate

Alexa Fluor® 647

Host

Alpaca

Reactivity

Mouse IgG1

Cat No : sms1AF647-1

Synonyms

mouse IgG1; mouse immunoglobulin Gamma 1

Validation Data Gallery

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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).

    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).

Product Information

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.

DescriptionNano-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.
ApplicationsIF, WB, FC
HostAlpaca
Species ReactivityMouse IgG1 Fc-fragment
No cross-reactivity: goat, guinea pig, rabbit, rat, sheep, human, macaque serum proteins, mouse IgG2a, IgG2b, IgG2c, IgG3
ConjugateAlexa Fluor® 647
Physical StateLiquid
Suggested DilutionImmunofluorescence 1:500
Super-resolution microscopy 1:500
Western blot 1:1,000
Purification MethodRecombinant expression, affinity purification IMAC
TypeMixture of 2 monoclonal Nanobodies
ClassRecombinant
Affinity (KD) CTK0103: KD =0.13 nM, CTK0104: KD = 0.63nM
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_2827579
NoteThis product is for research use only, not for diagnostic or therapeutic use

Publications

ApplicationTitle
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
View Article

Cell Rep

Hypothalamic astrocytic-BMAL1 regulates energy homeostasis in a sex-dependent manner

Authors - María Luengo-Mateos
View Article

PLoS Pathog

Peculiar transcriptional reprogramming with functional impairment of dendritic cells upon exposure to transformed HTLV-1-infected cells

Authors - Auriane Carcone
View Article