rabbit anti flag polyclonal antibody pab  (Proteintech)

Journal: Viruses

Article Title: The M2 Protein of the Influenza A Virus Interacts with PEX19 to Facilitate Virus Replication by Disrupting the Function of Peroxisome

doi: 10.3390/v16081309

Figure Lengend Snippet: IAV M2 protein interacts with host factor PEX19. ( a – c ) HEK293T cells were co-transfected with the indicated combination of plasmids expressing Myc-tagged PEX19 and Flag-tagged SC09 (H1N1) M2 ( a ), WSN (H1N1) M2 ( b ), or AH05 (H5N1) M2 ( c ). Cell lysates were subjected to immunoprecipitation with mouse anti-Flag or anti-Myc mAb, and the bound proteins were detected by Western blotting. ( d ) Schematic representation of differences in amino acid sequences of M2. ( e ) A549 cells were infected with the WSN (H1N1) virus at an MOI of 2 for 24 h. Cell lysates were immunoprecipitated with a mouse anti-PEX19 mAb, and the bound proteins were detected by Western blotting. ( f ) HEK293T cells were co-transfected with plasmids expressing Myc-PEX19 and GFP or GFP-tagged BM2. Cell lysates were immunoprecipitated with a mouse anti-GFP mAb, and the bound proteins were detected by Western blotting. ( g , h ) A549 cells were transfected individually or in combination with plasmids expressing Flag-tagged WSN (H1N1) M2 and Myc-tagged PEX19 ( g ) or were infected with the WSN (H1N1) virus at an MOI of 1 ( h ). Cells were immunostained with anti-M2 and anti-PEX19 antibodies 24 h after transfection or the indicated timepoints p.i., and were visualized by confocal microscopy. The 3D analysis of images was performed using Image J.

Article Snippet: The primary antibodies used in this study include rabbit anti-Flag polyclonal antibody (pAb) (F7425; Sigma-Aldrich, Saint Louis, MO, USA), mouse anti-Flag monoclonal antibody (mAb) (F3165; Sigma-Aldrich), rabbit anti-Myc pAb (C3965; Sigma-Aldrich), mouse anti-Myc mAb (M4439; Sigma-Aldrich), mouse anti-actin mAb (sc-47778; Santa Cruz, Dallas, TX, USA), rabbit anti-GAPDH pAb (10494-1-AP; Proteintech, Wuhan, China), rabbit anti-GFP pAb (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP mAb (ab1218; Abcam, Cambridge, MA, USA), rabbit anti-GST pAb (A00097; Genscript, Nanjing, China), mouse anti-GST mAb (A00865; Genscript), rabbit anti-V5 pAb (14440-1-AP; Proteintech), mouse anti-V5 mAb (A01724; Genscript), rabbit anti-M2 pAb (GTX125951; GeneTex, Irvine, CA, USA), mouse anti-M2 mAb (ab5416; Abcam), rabbit anti-PEX19 pAb (GTX110721; GeneTex), mouse anti-PEX19 mAb (GT554; GeneTex), and mouse anti-PMP70 mAb (ab211533; Abcam).

Techniques: Transfection, Expressing, Immunoprecipitation, Western Blot, Infection, Virus, Confocal Microscopy

Journal: Viruses

Article Title: The M2 Protein of the Influenza A Virus Interacts with PEX19 to Facilitate Virus Replication by Disrupting the Function of Peroxisome

doi: 10.3390/v16081309

Figure Lengend Snippet: The cytoplasmic tail domain of M2 and the C terminus of PEX19 are key regions of interactions. ( a – c ) HEK293T cells were co-transfected with the indicated combinations of plasmids. At 48 h post transfection, cell lysates were immunoprecipitated with a mouse anti-Myc mAb ( a , c ), or a mouse anti-Flag mAb ( b ), and the bound proteins were detected by Western blotting with a rabbit anti-GST pAb and a rabbit anti-Myc pAb ( a ) or with a rabbit anti-Flag pAb and a rabbit anti-Myc pAb ( b , c ).

Article Snippet: The primary antibodies used in this study include rabbit anti-Flag polyclonal antibody (pAb) (F7425; Sigma-Aldrich, Saint Louis, MO, USA), mouse anti-Flag monoclonal antibody (mAb) (F3165; Sigma-Aldrich), rabbit anti-Myc pAb (C3965; Sigma-Aldrich), mouse anti-Myc mAb (M4439; Sigma-Aldrich), mouse anti-actin mAb (sc-47778; Santa Cruz, Dallas, TX, USA), rabbit anti-GAPDH pAb (10494-1-AP; Proteintech, Wuhan, China), rabbit anti-GFP pAb (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP mAb (ab1218; Abcam, Cambridge, MA, USA), rabbit anti-GST pAb (A00097; Genscript, Nanjing, China), mouse anti-GST mAb (A00865; Genscript), rabbit anti-V5 pAb (14440-1-AP; Proteintech), mouse anti-V5 mAb (A01724; Genscript), rabbit anti-M2 pAb (GTX125951; GeneTex, Irvine, CA, USA), mouse anti-M2 mAb (ab5416; Abcam), rabbit anti-PEX19 pAb (GTX110721; GeneTex), mouse anti-PEX19 mAb (GT554; GeneTex), and mouse anti-PMP70 mAb (ab211533; Abcam).

Techniques: Transfection, Immunoprecipitation, Western Blot

Journal: Viruses

Article Title: The M2 Protein of the Influenza A Virus Interacts with PEX19 to Facilitate Virus Replication by Disrupting the Function of Peroxisome

doi: 10.3390/v16081309

Figure Lengend Snippet: The M2 protein disturbs the interactions between PEX19 and peroxisome-associated factors. ( a , b ) HEK293T cells were co-transfected with plasmids expressing Myc-PEX19, V5-PMP24 ( a ), or V5-PEX14 ( b ) and gradually increased amount of Flag-tagged WSN (H1N1) M2. At 24 h post transfection, cell lysates were immunoprecipitated with a mouse anti-Myc mAb, and the bound proteins were Western blotted with a rabbit anti-Flag, anti-Myc, or anti-V5 pAb.

Article Snippet: The primary antibodies used in this study include rabbit anti-Flag polyclonal antibody (pAb) (F7425; Sigma-Aldrich, Saint Louis, MO, USA), mouse anti-Flag monoclonal antibody (mAb) (F3165; Sigma-Aldrich), rabbit anti-Myc pAb (C3965; Sigma-Aldrich), mouse anti-Myc mAb (M4439; Sigma-Aldrich), mouse anti-actin mAb (sc-47778; Santa Cruz, Dallas, TX, USA), rabbit anti-GAPDH pAb (10494-1-AP; Proteintech, Wuhan, China), rabbit anti-GFP pAb (AG279; Beyotime Biotech, Shanghai, China), mouse anti-GFP mAb (ab1218; Abcam, Cambridge, MA, USA), rabbit anti-GST pAb (A00097; Genscript, Nanjing, China), mouse anti-GST mAb (A00865; Genscript), rabbit anti-V5 pAb (14440-1-AP; Proteintech), mouse anti-V5 mAb (A01724; Genscript), rabbit anti-M2 pAb (GTX125951; GeneTex, Irvine, CA, USA), mouse anti-M2 mAb (ab5416; Abcam), rabbit anti-PEX19 pAb (GTX110721; GeneTex), mouse anti-PEX19 mAb (GT554; GeneTex), and mouse anti-PMP70 mAb (ab211533; Abcam).

Techniques: Transfection, Expressing, Immunoprecipitation, Western Blot

Journal: Engineering Microbiology

Article Title: Identification of host proteins that interact with African swine fever virus pE301R

doi: 10.1016/j.engmic.2024.100149

Figure Lengend Snippet: Validation of Cap-host protein interactions. (A, B) HEK293T cells were co-transfected with a plasmid expressing FLAG-pE301R, together with plasmids expressing Myc-AIFM1 and Myc-VIM, respectively. Among them, FLAG-pE301R co-transfected with an empty vector served as the negative control. Cell lysates were immunoprecipitated with FLAG pAbs, or Myc pAbs and protein A/G beads, separated via SDS-PAGE, and then subjected to immunoblotting with the corresponding primary and secondary antibodies.

Article Snippet: Anti-FLAG mAbs (F1804) were obtained from Sigma-Aldrich (St. Louis, MO, USA) and anti-FLAG rabbit polyclonal Abs (pAbs) (A00170–40) were obtained from GenScript (Nanjing, China).

Techniques: Biomarker Discovery, Transfection, Plasmid Preparation, Expressing, Negative Control, Immunoprecipitation, SDS Page, Western Blot