Protocol detail

Antibodies for Chlamydia trachomatis study

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Polyclonal antibodies against C. trachomatis HtrA were generated previously (Huston et al., 2008 (link)). Monoclonal antibodies against EEA1 (610457, 1:100), SNX1 (611483,1:200) and SNX2 (611308, 1:200) were supplied by BD Bioscience. Monoclonal antibodies against the myc epiptope (9B11, 1:2000) were supplied by Abcam. Rabbit polyclonal antibodies against GFP (A-6455, 1:500) were purchased from Molecular Probes (Invitrogen). Rabbit polyclonal antibodies against Rab5 (C8B1, 1:100) were from Cell Signaling Technology. Goat polyclonal antibodies against Vps35 (IMG-3575, 1:400) were from Imgenex. Secondary antibodies were purchased from Molecular Probes (Life Technologies) and Li-Cor Bioscience. Wortmannin was supplied by Sigma-Aldrich (W1628). VPS34-In1 was from Merck Millipore (532628).

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Paul B., Kim H.S., Kerr M.C., Huston W.M., Teasdale R.D, & Collins B.M. (2017). Structural basis for the hijacking of endosomal sorting nexin proteins by Chlamydia trachomatis. eLife, 6, e22311.

Publication 2017

Monoclonal antibodies against (A-6455 Wortmannin VPS34-In1

Antibodies Goat Molecular probes Monoclonal antibodies Rabbit Vps34 in1 Wortmannin

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Variable analysis

independent variables

Wortmannin (supplied by Sigma-Aldrich (W1628))
VPS34-In1 (from Merck Millipore (532628))

dependent variables

Not explicitly mentioned

control variables

Polyclonal antibodies against C. trachomatis HtrA (generated previously)
Monoclonal antibodies against EEA1 (610457, 1:100), SNX1 (611483,1:200) and SNX2 (611308, 1:200) (supplied by BD Bioscience)
Monoclonal antibodies against the myc epiptope (9B11, 1:2000) (supplied by Abcam)
Rabbit polyclonal antibodies against GFP (A-6455, 1:500) (purchased from Molecular Probes (Invitrogen))
Rabbit polyclonal antibodies against Rab5 (C8B1, 1:100) (from Cell Signaling Technology)
Goat polyclonal antibodies against Vps35 (IMG-3575, 1:400) (from Imgenex)
Secondary antibodies (purchased from Molecular Probes (Life Technologies) and Li-Cor Bioscience)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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