Antibody Characterization for IFA and WB

The following antibodies were used for IFA and Western blotting at the indicated dilutions: rabbit polyclonal anti-HA SG77 (Thermo Fisher; 1:1,000 [WB]), mouse anti-PbHSP70 monoclonal antibody 4C9 (73 (link)) (1:1,000 [WB]), mouse anti-FLAG monoclonal antibody M2 (Sigma; 1:1,000 [IFA]), goat polyclonal anti-UIS4 (LS Biolabs LS-C204260; 1:1,000 [IFA]), and rabbit polyclonal anti-LISP2 (1:1,000 [IFA]). Anti-LISP2 antiserum was generated by GenScript against the peptide NGQKGNVDEERKSM\, located between the repeat region and 6-Cys domain, a portion of LISP2 previously shown to localize to the PV but not observed to enter the host cell (39 (link)). Purified antiserum labeled the parasite periphery in immunofluorescence assays, and loss of signal following EXP2 knockdown provided validation of specificity.

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Hussain T., Linera-Gonzalez J., Beck J.M., Fierro M.A., Mair G.R., Smith R.C, & Beck J.R. (2022). The PTEX Pore Component EXP2 Is Important for Intrahepatic Development during the Plasmodium Liver Stage. mBio, 13(6), e03096-22.

Publication 2022

anti-HA SG77

Anti antibody Antibodies Antiserum Assays immunofluorescence Cell Dilutions Goat Monoclonal antibody Mouse Parasite Peptide Rabbit

Corresponding Organization : Iowa State University

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Variable analysis

independent variables

Rabbit polyclonal anti-HA SG77 antibody (Thermo Fisher; 1:1,000 [WB])
Mouse anti-PbHSP70 monoclonal antibody 4C9 (1:1,000 [WB])
Mouse anti-FLAG monoclonal antibody M2 (Sigma; 1:1,000 [IFA])
Goat polyclonal anti-UIS4 antibody (LS Biolabs LS-C204260; 1:1,000 [IFA])
Rabbit polyclonal anti-LISP2 antibody (1:1,000 [IFA])

dependent variables

Immunofluorescence assay (IFA) labeling
Western blotting (WB) signal

control variables

Peptide used to generate anti-LISP2 antibody (NGQKGNVDEERKSM)
EXP2 knockdown experiments to validate anti-LISP2 antibody specificity

positive controls

Anti-LISP2 antiserum labeling the parasite periphery in immunofluorescence assays

negative controls

Loss of anti-LISP2 signal following EXP2 knockdown

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