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Rab11

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Rab11 is a small GTPase protein that plays a key role in regulating intracellular membrane trafficking, particularly in the recycling of endosomes. It is involved in the transport and sorting of various cargo proteins within the cell.

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Lab products found in correlation

Flag tag (flag), by Merck Group (2 mentions) Ab18061, by Abcam (1 mentions) Ab71333, by Abcam (1 mentions) H9658, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Ab150388, by Abcam (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) T6557, by Merck Group (1 mentions) Oregon green gelatin, by Thermo Fisher Scientific (1 mentions) Ezrin, by Abcam (1 mentions) Nherf1, by Abcam (1 mentions) Phalloidin 647, by Thermo Fisher Scientific (1 mentions) Hsp90 f 8, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Integrin α2, by Abcam (1 mentions) F actin, by Thermo Fisher Scientific (1 mentions) Nucblue fixed cell stain readyprobe, by Thermo Fisher Scientific (1 mentions) Proteostat, by Enzo Life Sciences (1 mentions) E cadherin, by Thermo Fisher Scientific (1 mentions) Nak atpase, by Thermo Fisher Scientific (1 mentions)

8 protocols using rab11

1

Immunofluorescence Analysis of Cell Adhesion

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Antibodies to paxillin (Abcam, ab2264, rabbit poly Ab), phalloidin-TRITC (Sigma, P1951), tubulin (Millipore, MAB1864, rat mAb), actinin (Abcam, ab18061, mouse mAb) talin (Abcam, ab71333, rabbit poly Ab), β1 integrin (Abcam, clone 12G10, ab30394, mouse mAb; and BD Biosciences, clone 9EG7, 553715, rat mAb), MB1.2 (Millipore, MAB1997, rat mAb), Rab5 (Cell Signaling Technology, 3547, rabbit mAb), Rab11 (Thermo Fisher Scientific, 71–5300, rabbit polyAb), Rab7 (Thermo Fisher Scientific, PA5–22959, rabbit polyAb), IQGAP1 (BD Biosciences, 610612, mouse mAb; and Abcam, ab86064, rabbit poly Ab), ARF6 (Cell Signaling Technology, 3546, rabbit poly Ab), HA (Sigma, H9658, mouse mAb), E-cadherin (Cell Signaling Technology, 3195, rabbit mAb), GAPDH-peroxidase; (Sigma, G9295, mouse mAb), EFA6 (Atlas Antibodies, HPA059237, rabbit poly Ab), EFA6 (Thermo Fisher Scientific, PA5–31153, rabbit poly Ab), and ARNO (Sigma, SAB2500109, goat poly Ab) were purchased, as stated for each above.
Ghosh M., Lo R., Ivic I., Aguilera B., Qendro V., Devarakonda C, & Shapiro L.H. (2019). CD13 tethers the IQGAP1-ARF6-EFA6 complex to the plasma membrane to promote ARF6 activation, B1 integrin recycling, and cell migration. Science signaling, 12(579), eaav5938.
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2

Immunoblotting and Immunofluorescence Protocols

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Antibodies used for immunoblotting were: TAO3 (ab150388, 1:1000, Abcam), CAMK1 (ab68234, 1:1000, Abcam), RAB11 (610657, 1:1000, BD Biosciences) and tubulin (T6557, 1:3000, Sigma).
Antibodies used for immunofluorescence staining were: TAO3 (LS-B11301, 1:250, LSBio), tubulin (6074, 1:250, Sigma) and RAB11 (71-5300, 1:250, Thermo Fisher Scientific). The antibodies for TKS5 (Rabbit monoclonal, F4 and mouse monoclonal, G6) were generated and validated by the Courtneidge laboratory, and will be fully described in a separate publication (in submission).
Phalloidin (Alexa fluor-350, -488, -568 or -647, Thermo Fisher Scientific) was used for actin staining. For the gelatin degradation assay, Oregon green gelatin (0.2 mg/ml in PBS containing 2% sucrose; Invitrogen) was used. Hoechst (1:4000, Thermo Fisher Scientific) was used for nuclear staining.
Iizuka S., Elia L., Navarro J.C., Gribbin K.P., Ardecky R., Abelman M.M., Ma C., Sergienko E., Zeng F., Pass I., Thomas G., McWeeney S.K., Hassig C.A., Pinkerton A.B., & Courtneidge S.A. (2020). The serine-threonine kinase TAO3 promotes cancer invasion and tumor growth by facilitating trafficking of endosomes containing the invadopodia scaffold TKS5α.
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3

Immunofluorescence Staining of Podocyte Markers

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Primary antibodies were diluted 1:100 unless stated otherwise: Endoglycan (R&D, AF3534, 1:200), Podocalyxin (R&D, MAB1556), Calbindin (Swant, CB-38a, 1:1000), NCAM (DSHB, 5B8; GeneTex H28-123), Rab11 (Invitrogen, 71-5300), Ezrin (Abcam, ab41672), CD34 (eBioscience, RAM34), Par-6b (SCBT, sc-67392, 1:400), Par-3 (Millipore, 07-330), aPKC (SCBT, C-20, 1:300), NHERF1 (Abcam, ab3452), Phalloidin 647 (Invitrogen, 42008A, 1:200). Secondary antibodies were purchased from Jackson ImmunoResearch, PA, USA.
Yang Z., Zimmerman S.E., Tsunezumi J., Braitsch C., Trent C., Bryant D.M., Cleaver O., González-Manchón C, & Marciano D.K. (2016). Role of CD34 family members in lumen formation in the developing kidney. Developmental biology, 418(1), 66-74.
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4

Cell Lysis and Western Blot Analysis

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Cells were lysed in IP buffer (20 mM Tris (pH 7.5), 150 mM NaCl, 2 mM EDTA, 1% TritonX-100, phenylmethylsulfonylfluoride (PMSF, 100 mM), complete protease inhibitor cocktail (PIC)) or RIPA buffer supplemented with PMSF and PIC, and analyzed by Western blot with primary antibodies against: DEF6 (H00050619-B01; Abnova), GAPDH (sc-365062; Santa Cruz Biotechnology), RAB11 (700184, Invitrogen), HSP90 (F-8, Santa Cruz). Uncropped scans and (where applicable) additional exposures of all immunoblots are shown in a separate Source Data file, with molecular weight markers indicated.
Serwas N.K., Hoeger B., Ardy R.C., Stulz S.V., Sui Z., Memaran N., Meeths M., Krolo A., Yüce Petronczki Ö., Pfajfer L., Hou T.Z., Halliday N., Santos-Valente E., Kalinichenko A., Kennedy A., Mace E.M., Mukherjee M., Tesi B., Schrempf A., Pickl W.F., Loizou J.I., Kain R., Bidmon-Fliegenschnee B., Schickel J.N., Glauzy S., Huemer J., Garncarz W., Salzer E., Pierides I., Bilic I., Thiel J., Priftakis P., Banerjee P.P., Förster-Waldl E., Medgyesi D., Huber W.D., Orange J.S., Meffre E., Sansom D.M., Bryceson Y.T., Altman A, & Boztug K. (2019). Human DEF6 deficiency underlies an immunodeficiency syndrome with systemic autoimmunity and aberrant CTLA-4 homeostasis. Nature Communications, 10, 3106.
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5

Comprehensive Immunofluorescence Assay Protocols

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For immunofluorescence, antibodies and the probes used were as follows: F-actin (phalloidin–Alexa Fluor 488, catalog A12379 or phalloidin–Alexa Fluor 647, catalog A22287, Invitrogen); nuclei (NucBlue Fixed Cell Stain Ready Probes, catalog R37606, Invitrogen); aggresome (ProteoStat, catalog ENZ-51038, Enzo Life Sciences Inc.); UNC45A (catalog sc-101493, 1:50, Santa Cruz Biotechnology Inc.); villin (catalog sc-58897, 1:50, Santa Cruz Biotechnology Inc.); DRA (catalog ab83545, 1:100, Abcam); Rab 11 (catalog 71-5300, 1:25, Invitrogen); NHE3 (catalog NBP-82574, 1:500, Novus Biologicals); MYO5B (catalog HPA069773, 1:400, Sigma-Aldrich); Epcam (catalog AF960, 1:100, R&D); CFTR (catalog ab59394, 1:100, abcam); NA/K-ATPase (catalog MA5-32184, 1:100, Thermo Fisher); integrin-α2 (catalog ab181548, 1:100, Abcam); and E-cadherin (catalog 13-700445, 1:100, Thermo Fisher). For STED, F-actin (phalloidin, phalloidin-ATTO 550, Invitrogen) was used. For Western blot, the following antibodies were used: actin (catalog 5125, 1:1000, Cell Signaling); FLAG (catalog F3165, 1:1000, Sigma-Aldrich); GAPDH (catalog 14C10, 1:1000, Cell Signaling Technology); HSP90 (catalog 4874, 1:1000, Cell Signaling Technology); UNC45A (catalog 171328, 1:500, Abcam); and MYO5B (catalog PA5-49519, 1:500, Thermo Fisher).
Duclaux-Loras R., Lebreton C., Berthelet J., Charbit-Henrion F., Nicolle O., Revenu des Courtils C., Waich S., Valovka T., Khiat A., Rabant M., Racine C., Guerrera I.C., Baptista J., Mahe M.M., Hess M.W., Durel B., Lefort N., Banal C., Parisot M., Talbotec C., Lacaille F., Ecochard-Dugelay E., Demir A.M., Vogel G.F., Faivre L., Rodrigues A., Fowler D., Janecke A.R., Müller T., Huber L.A., Rodrigues-Lima F., Ruemmele F.M., Uhlig H.H., Del Bene F., Michaux G., Cerf-Bensussan N, & Parlato M. (2022). UNC45A deficiency causes microvillus inclusion disease–like phenotype by impairing myosin VB–dependent apical trafficking. The Journal of Clinical Investigation, 132(10), e154997.
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6

Immunofluorescence Imaging of Centrosomal Proteins

Cited 3 times
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Antibodies used included: Mib1 (M5948; Sigma-Aldrich, St. Louis, MO), PCM1 (H262; Santa Cruz, Santa Cruz, CA), Cep131 (A301-415A; Bethyl Laboratories, Montgomery, TX), Cep90 (Kim and Rhee, 2011 (link)), Ki-67 (ab15580; Abcam, Cambridge, MA), GT335 (AG-20B-0020-C100; Adipogen, Switzerland), BBS4 (Kim et al., 2012 (link)), Rab11 (71–5300; Life Technologies, Carlsbad, CA), Rab8 (a gift from J. Peranen, University of Helsinki, Helsinki, Finland and 610844; BD Biosciences, San Jose, CA), α-tubulin (T5168; Sigma-Aldrich), Cep290 (A301-659A; Bethyl Laboratories), Ofd1 (a gift from J. Reiter, University of California, San Francisco, USA; Singla et al., 2010 (link)), Talpid3 (Kobayashi et al., 2014 (link)), Myc (sc-40; Santa Cruz), Flag (F3165; Sigma-Aldrich), centrin (04–1624; Millipore, Billerica, MA), GFP (G1544; Santa Cruz), Cep164 (a gift from Eva Lee, University of California, USA), IFT88 (13967-1-AP; Proteintech, Chicago, IL).
Wang L., Lee K., Malonis R., Sanchez I, & Dynlacht B.D. (2016). Tethering of an E3 ligase by PCM1 regulates the abundance of centrosomal KIAA0586/Talpid3 and promotes ciliogenesis. eLife, 5, e12950.
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7

Immunofluorescence Staining of Cellular Markers

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The following antibodies were used in this study: Pan cytokeratin AE1/AE3 (Abcam/ab27988) 1:50, Rab7 (Abcam/ab50533) 1:100, Rab11 (Life Technologies/715300) 1:100, GP135 (DSHB/3F2/D8) 1:100, Cingulin (Prekeris Lab) 1:100, FIP1 (Prekeris Lab) 1:200, FIP5 [Prekeris Lab (1:100)], Alexa 488 Anti-Rabbit secondary (Jackson ImmunoResearch/711-545-152) 1:200, Alexa 488 Anti-Mouse secondary (Jackson ImmunoResearch/715-545-150) 1:200, Alexa-568 Phalloidin (Invitrogen/A12380) 1:100.
Jewett C.E., Appel B.H, & Prekeris R. (2020). The Rab11 effectors Fip5 and Fip1 regulate zebrafish intestinal development. Biology Open, 9(10), bio055822.
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8

Localization of Rab11 in RBMVECs

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RBMVECs were grown on 10 mm cover slips in 4 well plates. All steps were performed at RT (room temperature) unless otherwise mentioned. After the 20 min treatment with conjugated VLPs the cells were washed with PBS, fixed and then permeabilized with 0.5% tritonX-100 in PBS at RT for 10 min. Following incubation with a blocking solution containing 5% goat serum and 0.3% tritonX-100 in PBS for 20 min, cells were incubated with primary antibodies Rab11 (Mouse Monoclonal Antibody- Life technologies) at 4 °C overnight, washed with PBS, and probed with the required RhoB-conjugated secondary antibody (Goat Anti-Mouse IgG (H + L) Antibody, Life Technologies). The treated coverslips were mounted onto a slide with Prolonged Gold antifade reagent with DAPI (Life Technologies). Fluorescence images were taken by using a Perkin Elmer UltraView ERS a spinning disk confocal microscope.
Anand P., O’Neil A., Lin E., Douglas T, & Holford M. (2015). Tailored delivery of analgesic ziconotide across a blood brain barrier model using viral nanocontainers. Scientific Reports, 5, 12497.
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