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3 protocols using donkey anti mouse igg conjugated to alexa fluor 680

1

Immunofluorescence Antibody Staining Protocol

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Primary antibodies were from the following sources: Mouse monoclonal antibodies against the peroxisomal membrane protein PMP70 and FLAG epitope from Sigma (St. Louis, MO, USA); mouse monoclonal against beta-actin, goat polyclonal antibody against GFP and rabbit polyclonal antibodies to PEX3, PEX7, PEX11B, PEX13, PEX19 and catalase from Abcam (Cambridge, MA, USA); rabbit polyclonal antibody to the tripeptide SKL was produced as described in [23 (link)]; goat polyclonal antibody to ZIKV NS5 was produced and purified as described in [31 (link)]; mouse monoclonal antibody against MAVS was from Santa Cruz Biotechnology (Santa Cruz, CA, USA); mouse monoclonal antibody against myc from Millipore (Burlington, MA, USA). Donkey anti-mouse IgG conjugated to Alexa Fluor 680, goat anti-rabbit IgG conjugated to Alexa Fluor 680, donkey anti-mouse IgG conjugated to Alexa Fluor 488, donkey anti-goat IgG conjugated to Alexa Fluor 680, chicken anti-goat IgG conjugated to Alexa Fluor 647, and donkey anti-rabbit IgG conjugated to Alexa Fluor 546 were purchased from Invitrogen (Carlsbad, CA, USA).
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2

Antibody Characterization for Peroxisomal Proteins

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Mouse monoclonal antibodies against the peroxisomal membrane protein PMP70 (Sigma, St. Louis, MO), HIV-1 p24 (Abcam, Cambridge, MA), GFP (Abcam), myc (Millipore Sigma, Etobicoke, ON, Canada), and beta-actin (Abcam) were purchased from the indicated suppliers. Rabbit polyclonal antibodies to PEX7, PEX11B, PEX13, PEX19, catalase, beta-catenin, and AU1 were obtained from Abcam; rabbit polyclonal antibody to PEX2 (PXMP3) was purchased from Pierce (Rockford, IL); rabbit monoclonal antibody to β-TrCP (D12C8) was purchased from New England Biolabs (Whitby, ON, Canada); and rabbit polyclonal antibody to the tripeptide SKL was produced as previously described (61 (link)).
Donkey anti-mouse IgG conjugated to Alexa Fluor 680, goat anti-rabbit IgG conjugated to Alexa Fluor 800, donkey anti-mouse IgG conjugated to Alexa Fluor 488, donkey anti-rabbit IgG conjugated to Alexa Fluor 488, donkey anti-rabbit IgG conjugated to Alexa Fluor 546, and donkey anti-mouse IgG conjugated to Alexa Fluor 546 were purchased from Invitrogen.
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3

Peroxisomal Protein Immunodetection

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Mouse monoclonal antibodies against the peroxisomal membrane protein PMP70 (Sigma, St. Louis, MO), HIV-1 p24 (Abcam, Cambridge, MA), and beta-actin (Abcam, Cambridge, MA) were purchased from indicated suppliers. Rabbit polyclonal antibodies to PEX7, PEX11B, PEX13, PEX19 and catalase were from Abcam (Cambridge, MA); Rabbit polyclonal antibody to PEX2 (PXMP3) was purchased from Pierce (Rockford, IL); Rabbit polyclonal antibody to thiolase (ACAA1) was from MyBioSource (San Diego, CA); Rabbit polyclonal antibody to the tri-peptide SKL were produced as previously described [60 (link)].
Donkey anti-mouse IgG conjugated to Alexa Fluor 680, goat anti-rabbit IgG conjugated to Alexa Fluor 680, donkey anti-mouse IgG conjugated to Alexa Fluor 488, donkey anti-rabbit IgG conjugated to Alexa Fluor 488, and donkey anti-mouse IgG conjugated to Alexa Fluor 546 were purchased from Invitrogen (Carlsbad, CA).
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