Anti rab11

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Anti-Rab11 is a laboratory reagent used to detect and quantify the Rab11 protein in biological samples. Rab11 is a member of the Ras-related protein family and is involved in the regulation of intracellular membrane trafficking. Anti-Rab11 can be used in various research applications, including immunohistochemistry, Western blotting, and immunoprecipitation, to study the localization and expression of Rab11 in cells and tissues.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Rab11 (8 citations) Rabbit anti-Rab11 (4 citations)

9 protocols using anti rab11

Extracellular Vesicle Characterization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies: anti-Cyclophilin A (1:1000 dilution, Abcam, ab126738), anti-CD147 (1:1000, ThermoFisher, MA1–19,201), anti-HSP70 (1:3000, Santa Cruz, sc-66,048), anti-TSG101 (1:1000 dilution, Abcam, ab125011), anti-HSP90B1 (1:1000 dilution, Cusabio, CSB-PA10887A0Rb), anti-GAPDH (1:500 dilution, EMD Millipore, MAB374), anti-Rab7 (1:1000 dilution, Cell signaling Technology, 9367), anti-Rab11 (1:250 dilution, ThermoFisher, 71–5300).
The secondary antibodies: anti-Rabbit IgG-DyLight 800 (1:10,000 dilution, ThermoFisher, SA5–35,571), anti-Mouse IgG-DyLight 680 (1:10,000 dilution, ThermoFisher, 35,519)
Reagents: Trypsin (Pierce Trypsin Protease, MS Grade, ThermoFisher, 90,057), Propidium Iodide (ThermoFisher, P1304MP), 5-(and −6)-Carboxyfluorescein Diacetate, Succinimidyl Ester, mixed isomers (5(6)-CFDA, SE) (ThermoFisher, C1157)

Wu Y., Brennan K., Fernández A.B, & Mc Gee M.M. (2021). Cyclophilin A regulates secretion of tumour-derived extracellular vesicles. Translational Oncology, 14(8), 101112.

+ Open protocol

+ Expand

Comprehensive Protein Immunodetection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following primary antibodies were used: Anti-Arfgef1 N-terminal epitope (ThermoFisher Scientific, PA5-54894; Santa Cruz, sc-376790; sc-50391), anti-Arfgef1 C-terminal epitope (Bethyl, A300-998A), anti-Arfgef2 C-terminal epitope (Sigma-Aldrich, MABS1246), Anti-GABA_A Receptor with epitope for extracellular domain of the β2,3 subunits (Millipore Sigma, MAB341), anti-synaptophysin (ThermoFisher Scientific, PA1-1043), anti-lamp1 (DSHB, 1D4B), anti-eea1 (R&D Systems, AF8047), anti-transferrin receptor (Abcam, ab84036), anti-Rab11 (ThermoFisher Scientific, 71-5300), anti-Arf6 (ThermoFisher Scientific, PA1-093), anti-map2 (Millipore Sigma, Ab5622), anti-ankG (Santa Cruz, sc-28561), anti-ß-tubulin (Proteintech, 10094-1-AP and 66240-1-Ig), anti-GAPDH (Proteintech, 60004-1-Ig), anti-lamin B1 (Proteintech, 66095-1-lg). The following secondary antibodies were used: Goat anti-mouse HRP-conjugated IgG (Proteintech, SA00001-1), goat anti-rabbit HRP-conjugated IgG (Proteintech, SA00001-2), Goat anti-mouse Alexa Fluor 488-conjugated IgG (ThermoFisher Scientific, A11017), goat anti-rabbit Alexa Fluor 594-conjugated IgG (ThermoFisher Scientific, A11072), goat anti-rat Alexa Fluor 594-conjugated IgG (ThermoFisher Scientific, A11007), donkey anti-sheep NL557-conjugated IgG (R&D Systems, NL010).

Teoh J., Subramanian N., Pero M.E., Bartolini F., Amador A., Kanber A., Williams D., Petri S., Yang M., Allen A.S., Beal J., Haut S.R, & Frankel W.N. (2019). Arfgef1 haploinsufficiency in mice alters neuronal endosome composition and decreases membrane surface postsynaptic GABAA receptors. Neurobiology of disease, 134, 104632.

+ Open protocol

+ Expand

Antibody Source and Reagent Acquisition

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-actin (A2066) was purchased from Sigma-Aldrich (St. Louis, MS). Anti-synapsin I (Ab64581) was obtained from abcam (Tokyo, Japan). Anti-PSD-95 (MA1-045) and anti-Rab11 were from Thermo Fisher Scientific (Grand Island, NY). Anti-p35 (C-19) and anti-Cdk5 (DC-17) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-GluR2/3 was from Millipore (Burlington, MA). Anti-NR2A antibody was purchased from Chemicon (Temecula, CA). Anti-VAMP-2 was provided by Dr. M. Itakura at Kitasato University. Anti-LMTK1 was generated by immunizing rabbits with the LMTK1 peptide of amino acids 651–853 (Fig. 1B). AlexaFluor 488-conjugated anti-mouse (AB_2556548) and AlexaFluor 568-conjugated anti-rabbit IgG (AB_143157) were obtained from Thermo Fisher Scientific. HRP-conjugated anti-mouse (AB_2617137) and anti-rabbit antibodies (AB_2617138) were from Agilent Dako (Santa Clara, CA). Picrotoxin and methylphenidate were purchased from Sigma-Aldrich. d-(-)-2-amino-5-phosphonovaleric acid (D-APV) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were purchased from Tocris Bioscience (Avonmouth, UK).

Takahashi M., Sugiyama A., Wei R., Kobayashi S., Fukuda K., Nishino H., Takahashi R., Tsutsumi K., Kita I., Ando K., Manabe T., Kamiguchi H., Tomomura M, & Hisanaga S.I. (2020). Hyperactive and impulsive behaviors of LMTK1 knockout mice. Scientific Reports, 10, 15461.

+ Open protocol

+ Expand

Visualizing Rab11, EEA-1, and Lamp-1 Localization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hela cells, which expressed EGFP-SH3BP5 or EGFP-SH3BP5L transiently, were cultured on coverslips and then washed with PBS twice. The cells were fixed with 4% paraformaldehyde in PBS for 15 min and washed three times. Then, we permeabilized cells with 0.05% saponin and incubated them with PBS containing 5% FBS for 30 min for blocking and treated with the following antibodies. We used anti-Rab11 (1:250 dilution; #71-5300; Invitrogen), anti-EEA-1 (1:250 dilution; #610456; BD Biosciences), and anti-Lamp-1 (1:250 dilution; #sc-20011; Santa Cruz Biotechnology) antibodies as the primary antibodies and goat anti-rabbit or anti-mouse Alexa Fluor 594 (1:1,000; Life Technologies Inc.). We acquired images using an FV1200 confocal microscope (Olympus) with a 100× PlanApo oil immersion lens (1.40 numerical aperture; Olympus). For mitochondria staining, we incubated Hela cells, which expressed EGFP-SH3BP5 or EGFP-SH3BP5L transiently in glass bottom dishes, with a MitoTracker Red CMXRos (0.1 nM; M7512; Invitrogen) for 15 min and washed them with PBS twice for observation.

Goto-Ito S., Morooka N., Yamagata A., Sato Y., Sato K, & Fukai S. (2019). Structural basis of guanine nucleotide exchange for Rab11 by SH3BP5. Life Science Alliance, 2(2), e201900297.

+ Open protocol

+ Expand

Indirect Immunofluorescence Microscopy Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For indirect immunofluorescence assays cells were grown on glass coverslips, fixed with PBS-4% PFA, and permeabilized with PBS-0.1% Triton X-100 or with PBS-0.25% Saponin. The cells were incubated with the anti-NP (Meridian AA5H, diluted 1/500), anti-Rab11 (Invitrogen 71-5300, diluted 1/50), and/or anti-PDI antibodies (Sigma P7372, diluted 1/200), and then with DyeLight550-coupled anti-rabbit and/or DyeLight488 anti-mouse IgG secondary antibodies (Pierce, diluted 1/500) and/or AF488-coupled anti-HA-tag antibody (Molecular Probes, diluted 1/250) or AF488-coupled anti-M2 antibody (Santa Cruz sc-32238, diluted 1/100). The samples were stained with Hoechst 33342 (Invitrogen, diluted to 1 μg/ml), mounted in ProLong Gold mounting medium (Life technologies P36930). For epifluorescence microscopy, images were acquired with upright Zeiss Axio Imager Z2 with HXP 120 lamp (at 50% level) using ×63 oil immersion objective lenses. Images were acquired with an AxioCam MRm camera and the ZEN blue 2012 software (Zeiss). For confocal microscopy images were acquired with a Leica TCS SP5 confocal multispectral microscope using a HCX PL APO 63.0 X/1.4 N.A. oil objective and LAS AF v.2.7.3 software (Leica Microsystems).

de Castro Martin I.F., Fournier G., Sachse M., Pizarro-Cerda J., Risco C, & Naffakh N. (2017). Influenza virus genome reaches the plasma membrane via a modified endoplasmic reticulum and Rab11-dependent vesicles. Nature Communications, 8, 1396.

+ Open protocol

+ Expand

Immunofluorescence staining of fixed cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were fixed with 3.7% formaldehyde 20 mM HEPES in PBS for 10min. This step was skipped when working with Jurkat cells as they were fixed during the setup of the cells. The formaldehyde was quenched with 10 mM Tris in PBS for 10mins, and the cells were then permeabilised for 5mins with PBS containing 0.05% Tx-100. The cells were then blocked with 3% BSA in PBS for 20mins and then stained with primary antibodies (Ticilimumab, anti-LRBA (Atlas Antibodies), anti-Rab5, anti-Rab7, anti-Rab9 (all Cell Signaling Technology), anti-Rab11 (Invitrogen)) in 3mg/ml BSA in PBS for 1hr at RT. Next, goat anti-human IgG-AlexaFluor 546, donkey anti-rabbit IgG-AlexaFluor 488 (both Invitrogen), Hoechst 33342 (Thermo Fisher Scientific) and 2.5 μM CellTrace Violet (Molecular Probes) in 3mg/ml BSA in PBS were added to the cells for 45mins. All steps up to and including the 3% BSA block were carried out on ice, all the following steps were at RT, and the cells were washed 3 times with PBS between each step.

Janman D., Hinze C., Kennedy A., Halliday N., Waters E., Williams C., Rowshanravan B., Hou T.Z., Minogue S., Qureshi O.S, & Sansom D.M. (2021). Regulation of CTLA‐4 recycling by LRBA and Rab11. Immunology, 164(1), 106-119.

+ Open protocol

+ Expand

Immunoblotting Antibody Characterization

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-p35 (C-19), anti-Cdk5 (DC17), and monoclonal anti-myc (9E10) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-Rab11 was from Invitrogen (Carlsbad, CA). Monoclonal anti–neuron-specific class III β-tubulin was from Techne Corporation (Minneapolis, MN). Anti-actin was from Sigma-Aldrich (Saint Louis, MO). Anti-PSD95 was from Thermo Scientific (Manor Park Runcom, Cheshire, United Kingdom). Anti-GluR2 was from Chemicon (Temecula, CA). Anti-LMTK1 and anti–phospho-Ser34 of LMTK1 (pS34) have been described (Tsutsumi et al., 2008 (link), 2010 (link)). Alexa-conjugated secondary antibodies were from Invitrogen.

Takano T., Urushibara T., Yoshioka N., Saito T., Fukuda M., Tomomura M, & Hisanaga S.I. (2014). LMTK1 regulates dendritic formation by regulating movement of Rab11A-positive endosomes. Molecular Biology of the Cell, 25(11), 1755-1768.

+ Open protocol

+ Expand

Multicolor Immunofluorescence Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were fixed with 3.7% formaldehyde 20 mM HEPES in PBS for 10 min. This step was skipped when working with Jurkat cells as they were fixed during the set‐up of the cells. The formaldehyde was quenched with 10 mM Tris in PBS for 10 min, and the cells were then permeabilized for 5 min with PBS containing 0.05% Triton‐X‐100. The cells were then blocked with 3% BSA in PBS for 20 min and then stained with primary antibodies (ticilimumab and anti‐LRBA (Atlas Antibodies); anti‐Rab5, anti‐Rab7 and anti‐Rab9 (all from Cell Signaling Technology); anti‐Rab11 (Invitrogen or Atlas Antibodies) and anti‐LC3A/B (Cell Signaling Technology)) in 3 mg/ml BSA in PBS for 1 h at RT. Next, goat anti‐human IgG‐Alexa Fluor 546, donkey anti‐rabbit IgG‐Alexa Fluor 488 (both Invitrogen), Hoechst 33342 (Thermo Fisher Scientific) and 2.5 µM CellTrace Violet (Molecular Probes) in 3 mg/ml BSA in PBS were added to the cells for 45 min. All steps up to and including the 3% BSA block were carried out on ice, all the following steps were at RT, and the cells were washed 3 times with PBS between each step.

+ Open protocol

+ Expand

Antibody Labeling and Protein Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-myc (4A6) was purchased from Millipore (Billerica, MA, USA), anti-GFP from Roche Diagnostics (Manheim, Germany), anti-Rab11 from Invitrogen (Carlsbad, CA, USA), anti-b-tubulin, (TUB 2.1) and anti-c-tubulin (GTU-88) from Sigma-Aldrich (St. Louis, MO, USA), anti-transferrin receptor from Zymed Laboratories (South San Francisco, CA, USA) and anti-Golgin-97 and Alexa-conjugated secondary antibodies from Invitrogen. The anti-LMTK1A antibodies have been described previously (Tsutsumi et al. 2008 (Tsutsumi et al. , 2010)) . Tetramethylrhodamine isothiocyanate (TRITC) phalloidin was purchased from Sigma-Aldrich. HRP-conjugated anti-rabbit IgG and HRP-conjugated anti-mouse IgG were purchased from DAKO (Glostrup, Denmark). 4-(2-Aminoethyl)-benzenesulfonylfluoride hydrochloride (AEBSF) was purchased from Sigma-Aldrich, leupeptin from Peptide Institute (Osaka, Japan), DAPI from Dojindo (Kumamoto, Japan) and Protein-G Sepharose from GE Healthcare Bio-Science (Uppsala, Sweden).

Sharma G., Tsutsumi K., Saito T., Asada A., Ando K., Tomomura M, & Hisanaga S.I. (2016). Kinase activity of endosomal kinase LMTK1A regulates its cellular localization and interactions with cytoskeletons. Genes to cells : devoted to molecular & cellular mechanisms, 21(10).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!