Ha clone c29f4

Intracellular proteins were detected in permeabilized MDMs by flow cytometry with Alexa Fluor 488-, phycoerythrin-, Alexa Fluor 647- or biotin-labeled antibodies to phospho-ERK (clone E10), phospho-p38 (clone 3D7), phospho-JNK (clone G9), phospho-IκBα (clone 14D4), HA (clone C29F4), Myc (clone 9B11) (Cell Signaling Technology, Danvers, MA) or INAVA (Abcam, Cambridge, MA).

In-house rabbit polyclonal CatSper1 (Ren et al., 2001 (link)), CatSper3 (Qi et al., 2007 (link)), CatSperε (Chung et al., 2017 (link)) antibodies were described previously. Polyclonal CA-IV antibody (M-50) was purchased from Santacruz. Monoclonal antibodies were purchased from BD Biosciences: anti-caveolin1 (clone 2297); EMD Milipore: anti-phosphotyrosine (clon4G10), anti-acetylated tubulin (clone 6-11B-1), anti-HA agarose (clone HA-7); Thermo Scientific: anti-HA magnetic beads; and Cell Signaling Technology: β-actin (clone 13E5) and HA (clone C29F4). HRP-conjugated goat anti-rabbit IgG and goat anti-mouse IgG were from Jackson Immunoresearch. PNA-Alexa 568, WGA-Alexa 555, WGA-Alexa 647, goat anti-mouse IgG (Alexa 488 or 647), and goat anti-rabbit IgG (Alexa 568 or Alexa 647) were from Invitrogen. H89, calyculin A, and calpain inhibitor I were purchased from Calbiochem. ST-Ht31 was from Promega. Calpain inhibitor II and III were from Enzo life science. All other chemicals were from Sigma-Aldrich unless indicated.