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Rabbit anti rab5

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Rabbit anti-Rab5 is an antibody raised in rabbits against the Rab5 protein. Rab5 is a small GTPase that plays a key role in early endocytic trafficking. The antibody can be used to detect and study the Rab5 protein.

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Lab products found in correlation

Lsm 710 confocal microscope, by Zeiss (3 mentions) Goat anti mouse, by Cell Signaling Technology (2 mentions) Pvegfr2 y1059, by Cell Signaling Technology (2 mentions) Pvegfr2 y1059, by Merck Group (2 mentions) Pvegfr2 y951, by Cell Signaling Technology (2 mentions) Total vegfr2, by Cell Signaling Technology (2 mentions) Ve cad, by Santa Cruz Biotechnology (2 mentions) Sc 9989, by BD (2 mentions) Ve cad, by BD (2 mentions) Sc 1718 r, by BD (2 mentions) Goat anti vegfr2, by R&D Systems (2 mentions) Rabbit anti ve cadherin, by Cell Signaling Technology (2 mentions) Pvegfr2 y1175, by Cell Signaling Technology (2 mentions) Rabbit anti ha, by Cell Signaling Technology (2 mentions) Ha tag, by Cell Signaling Technology (2 mentions) Mouse anti ha 11, by Fortrea (2 mentions) Rabbit anti rab7, by Cell Signaling Technology (2 mentions) Rabbit anti rab7 antibody, by Cell Signaling Technology (2 mentions) Anti rabbit dylight 649 antibody, by BioLegend (2 mentions) Anti v5 fitc antibody, by Thermo Fisher Scientific (2 mentions)

8 protocols using rabbit anti rab5

1

Western Blot and Flow Cytometry Analysis of Hepatitis B Virus Antigen and ER Proteins

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For western blot analysis, HBs antigen and calnexin were detected with goat anti-HBV polyclonal antibody (Murex, DiaSorin) coupled to horseradish peroxidase (HRP) and rabbit calnexin polyclonal antibody (Enzo), respectively. The rabbit anti-ERp46 (Abcam), mouse anti-ERp57 (Abcam), and mouse anti-ERp72 (Santa Cruz Biotechnology) antibodies were used for detecting PDI proteins by flow cytometry and western blot. NTCP was detected with polyclonal NTCP/SLC10A1 antibody (Bioss Antibodies) coupled to Phycoerythrin (PE) for flow cytometry. mouse anti-ERp57 (Abcam), Rabbit anti-Rab5, anti-Rab7, anti-Rab11, and anti-Lamp1 (Cell Signaling Technology), and Donkey anti-Rabbit-Alexa-Fluor-488 and Donkey anti-Mouse-Alexa-Fluor-568 (Thermo Fisher) antibodies were used for immunofluorescence (IF) studies.
Pérez-Vargas J., Teppa E., Amirache F., Boson B., Pereira de Oliveira R., Combet C., Böckmann A., Fusil F., Freitas N., Carbone A, & Cosset F.L. (2021). A fusion peptide in preS1 and the human protein disulfide isomerase ERp57 are involved in hepatitis B virus membrane fusion process. eLife, 10, e64507.
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2

Antibody Panel for Vascular Signaling Analysis

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List of antibodies with application and dilutions (IHC, immunohistochemistry; WB; western blot): pVEGFR2 Y1175 (WB 1:1000, Cell Signaling #2478), pVEGFR2 Y1059 (for HUVEC, WB 1:1000, Cell Signaling #3817, ), pVEGFR2 Y1059 (for mouse ECs, WB 1:1000, EMD Millipore #ABS553), pVEGFR2 Y951 (WB 1:1000, Cell Signaling #4991 ), VEGFR2 total (WB 1:1000, Cell Signaling #2479), VE-Cad (for HUVEC, WB 1:200, Santa Cruz #sc-9989 (F-8) ), VE-Cad (for mouse ECs, WB 1:500, BD Pharmingen #555289), mouse Sdc2 (WB 1:200, R&D #AF6585 - polyclonal raised against mouse Sdc2 ED), human Sdc2 (WB 1:200, R&D # AF2965, polyclonal raised against mouse human ED), mouse PTP1b (WB 1:200, Santa Cruz #sc-1718-R (N19-R)), human PTP1b (WB 1:500, BD Pharmingen #610139), VEPTP ( VE-PTP-C pAb69 and VE-PTP 1–8 pAb70 were gifts from Prof. Dietmar Vestweber, Max Plank Institute for Molecular Biomedicine, Münster, Germany), DEP1 (WB 1:200, Santa Cruz #sc-21761 (143–41)) NRP1 (WB 1:1000, Cell signaling #3725,), HA-tag (WB 1:1000, Cell signaling #3724), rabbit anti-Rab5 (Cell Signaling, #3547, IHC 1:200); goat anti-VEGFR2 (R&D, #AF357, IHC 1:100); mouse anti-HA.11 (Covance, #MMS-101P, IHC 1:200); rabbit anti-HA (Cell signaling, #3724, IHC 1:200); rabbit anti-VE-Cadherin (Cell Signaling, #2500, IHC 1:200); goat anti-mouse VE-Cadherin (R&D Systems, #AF1002, IHC 1:100); rat anti-mouse Flk1 (BD, #555307, IHC 1:100).
Corti F., Ristori E., Rivera-Molina F., Toomre D., Zhang J., Mihailovic J., Zhuang Z, & Simons M. (2022). Syndecan-2 selectively regulates VEGF-induced vascular permeability. Nature cardiovascular research, 1(5), 518-528.
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3

Visualizing Ciliary Protein Dynamics

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Approximately 0.6 × 105 cells per well were seeded in Lab-Tek
chambered slides and cultured for 24 hr. The cells were transfected, allowed to
recover for 24 to 36 hr, and then treated with ShhN-CM or other compounds, as
indicated. For visualizing ciliary proteins, the transfected cells were starved
in DMEM containing 0.5% FBS for 24 hr before other treatments. The cells were
fixed with 4% paraformaldehyde for 10 min at 4°C, and standard procedures
for immunostaining were followed. The primary antibodies used were rabbit
anti-Caveolin-1 (1:1000; Sigma-Aldrich (St. Louis, MO)), rabbit anti-Clathrin
heavy chain (1:200; Cell Signaling Technology (Danvers, MA)), rabbit anti-Rab5
(1:150, Cell Signaling Technology), rabbit anti-Rab7 (1:50, Cell Signaling
Technology), rabbit anti-Lamp1 (1:150; Sigma), mouse anti-acetylated Tubulin
(1:2000; Sigma), rabbit anti-Gli3 (1:500; R&D (Minneapolis, MN)), and
rabbit anti-Smo (1:500; a gift from Dr Rajat Rohatgi). Alexa-coupled secondary
antibodies were purchased from Life Technologies Corp.
Yue S., Tang L.Y., Tang Y., Tang Y., Shen Q.H., Ding J., Chen Y., Zhang Z., Yu T.T., Zhang Y.E, & Cheng S.Y. (2014). Requirement of Smurf-mediated endocytosis of Patched1 in sonic hedgehog signal reception. eLife, 3, e02555.
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4

Visualizing IFITM3 and Mycobacterium Trafficking

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A549 cells stably transduced with the IFITM3-V5 tag construct or the empty vector were seeded on a small coverslip in 12-well plates in fresh media and were infected with MTb-mCherry for 3 hr, after which the cultures were washed and fresh media were added. Cells were further incubated at 37°C for 20 hr. MTb-infected cells were fixed with 4% paraformaldehyde before they were blocked and permeabilized with buffer containing 5% normal donkey serum and 0.3% Triton X-100 in PBS for 2 hr. Cells were labeled with 1:150 rabbit anti-Rab5 or 1:100 rabbit anti-Rab7 antibody (Cell Signaling Technology) overnight in antibody dilution buffer containing 1% BSA followed by 1:500 secondary donkey anti-rabbit DyLight 649 antibody (BioLegend) for 2 hr. Cells were further labeled with mouse fluorescein isothiocyanate (FITC) anti-V5 antibody (Invitrogen) overnight, after which slides were mounted in mounting media containing DAPI. Images were captured with an Olympus (FV1000) confocal microscope and Fluoview Software 2. Analysis was performed using ImageJ software.
Ranjbar S., Haridas V., Jasenosky L.D., Falvo J.V, & Goldfeld A.E. (2015). A Role for IFITM Proteins in Restriction of Mycobacterium tuberculosis Infection. Cell reports, 13(5), 874-883.
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5

Antibody Panel for Vascular Signaling Analysis

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List of antibodies with application and dilutions (IHC, immunohistochemistry; WB; western blot): pVEGFR2 Y1175 (WB 1:1000, Cell Signaling #2478), pVEGFR2 Y1059 (for HUVEC, WB 1:1000, Cell Signaling #3817, ), pVEGFR2 Y1059 (for mouse ECs, WB 1:1000, EMD Millipore #ABS553), pVEGFR2 Y951 (WB 1:1000, Cell Signaling #4991 ), VEGFR2 total (WB 1:1000, Cell Signaling #2479), VE-Cad (for HUVEC, WB 1:200, Santa Cruz #sc-9989 (F-8) ), VE-Cad (for mouse ECs, WB 1:500, BD Pharmingen #555289), mouse Sdc2 (WB 1:200, R&D #AF6585 - polyclonal raised against mouse Sdc2 ED), human Sdc2 (WB 1:200, R&D # AF2965, polyclonal raised against mouse human ED), mouse PTP1b (WB 1:200, Santa Cruz #sc-1718-R (N19-R)), human PTP1b (WB 1:500, BD Pharmingen #610139), VEPTP ( VE-PTP-C pAb69 and VE-PTP 1–8 pAb70 were gifts from Prof. Dietmar Vestweber, Max Plank Institute for Molecular Biomedicine, Münster, Germany), DEP1 (WB 1:200, Santa Cruz #sc-21761 (143–41)) NRP1 (WB 1:1000, Cell signaling #3725,), HA-tag (WB 1:1000, Cell signaling #3724), rabbit anti-Rab5 (Cell Signaling, #3547, IHC 1:200); goat anti-VEGFR2 (R&D, #AF357, IHC 1:100); mouse anti-HA.11 (Covance, #MMS-101P, IHC 1:200); rabbit anti-HA (Cell signaling, #3724, IHC 1:200); rabbit anti-VE-Cadherin (Cell Signaling, #2500, IHC 1:200); goat anti-mouse VE-Cadherin (R&D Systems, #AF1002, IHC 1:100); rat anti-mouse Flk1 (BD, #555307, IHC 1:100).
Corti F., Ristori E., Rivera-Molina F., Toomre D., Zhang J., Mihailovic J., Zhuang Z, & Simons M. (2022). Syndecan-2 selectively regulates VEGF-induced vascular permeability. Nature cardiovascular research, 1(5), 518-528.
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6

Visualizing IFITM3 and Mycobacterium Trafficking

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A549 cells stably transduced with the IFITM3-V5 tag construct or the empty vector were seeded on a small coverslip in 12-well plates in fresh media and were infected with MTb-mCherry for 3 hr, after which the cultures were washed and fresh media were added. Cells were further incubated at 37°C for 20 hr. MTb-infected cells were fixed with 4% paraformaldehyde before they were blocked and permeabilized with buffer containing 5% normal donkey serum and 0.3% Triton X-100 in PBS for 2 hr. Cells were labeled with 1:150 rabbit anti-Rab5 or 1:100 rabbit anti-Rab7 antibody (Cell Signaling Technology) overnight in antibody dilution buffer containing 1% BSA followed by 1:500 secondary donkey anti-rabbit DyLight 649 antibody (BioLegend) for 2 hr. Cells were further labeled with mouse fluorescein isothiocyanate (FITC) anti-V5 antibody (Invitrogen) overnight, after which slides were mounted in mounting media containing DAPI. Images were captured with an Olympus (FV1000) confocal microscope and Fluoview Software 2. Analysis was performed using ImageJ software.
Ranjbar S., Haridas V., Jasenosky L.D., Falvo J.V, & Goldfeld A.E. (2015). A Role for IFITM Proteins in Restriction of Mycobacterium tuberculosis Infection. Cell reports, 13(5), 874-883.
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7

Immunostaining and Electron Microscopy Analysis

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For immunostaining, hematoxylin & eosin (H&E) and Periodic Acid Schiff (PAS) analysis, biopsies were fixed in 4% paraformaldehyde at room temperature for 30 m followed by storage in 70% ethanol until paraffin embedding. Sections were stained using the antibodies against the following proteins: mouse anti-Actin (RRID:AB_11004139, Invitrogen/Thermo-Fisher), rabbit anti-Zo1 (RRID:AB_2533938, Invitrogen), mouse anti-EPCaM (RRID:AB_10981962, Thermo Fisher, Waltham, MA), rabbit anti-rab5 (RRID:AB_823625, Cell Signaling Technologies), goat anti-Muc2 (RRID:AB_2146667, Santa Cruz Biotechnology, Dallas, TX), mouse anti-Salmonella (RRID:AB_1125358, BD Biosciences, San Jose, CA), rabbit-anti-Salmonella (RRID:AB_561201, Pierce/Thermo-Fisher), rabbit-anti-Salmonella-biotin conjugated (RRID:AB_1018415, Invitrogen/Thermo-Fisher) and mouse anti-tubulin (RRID:AB_2715541, Cell Signaling Technologies). Fluorescent, conjugated, secondary monoclonal antibodies were used for detection. For samples using streptavidin-biotin detection, avidin-biotin blocking was performed prior to staining (Life Technologies/Thermo-Fisher). Samples were imaged using a Nikon A1SiR confocal microscope.
For transmission electron microscopy (TEM) analysis, samples were fixed in 2%PFA/2.5% Glut in 0.1 M Sodium Cacodylate followed by mounting on grids and imaged using a transmission electron microscope (JEOL, Peabody, MA).
Nickerson K.P., Senger S., Zhang Y., Lima R., Patel S., Ingano L., Flavahan W.A., Kumar D.K., Fraser C.M., Faherty C.S., Sztein M.B., Fiorentino M, & Fasano A. (2018). Salmonella Typhi Colonization Provokes Extensive Transcriptional Changes Aimed at Evading Host Mucosal Immune Defense During Early Infection of Human Intestinal Tissue. EBioMedicine, 31, 92-109.
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8

Immunofluorescence staining of cells and tissues

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Tissues were fixed in Zn-formalin and embedded in paraffin prior to staining. Sections were deparaffinized, rehydrated and subjected to antigen retrieval. For staining cell lines, cells were fixed in 4% paraformaldehyde for 15 mins. For staining, sections and fixed cells were blocked in 5% donkey serum for 1 hour at room temperature (RT), incubated with primary antibodies for 1 hour at RT, washed, incubated with secondary antibodies for 1 hour at RT, washed and mounted. Primary antibodies used include goat anti-GFP (Abcam), rat anti-Ecadherin (Takara Bio), rabbit anti-Zeb1 (Santa Cruz), rabbit anti-Slug (gift of Dr. Joel Habener), rabbit anti-Vimentin (Cell Signaling Technologies), rabbit anti-Fsp1 (DAKO), rabbit anti-Rab5 (Cell Signaling Technologies), rabbit anti-Rab7 (Cell Signaling Technologies), rabbit anti-Rab11 (Cell Signaling Technologies), rabbit anti-EpCAM (Abcam), rabbit anti-Claudin-7 (Abcam), and rabbit anti-B-catenin (Cell Signaling Technologies). Zeb1 required additional tyramide signaling amplification (PerkinElmer). Slides were visualized using an Olympus IX71 inverted multicolor fluorescent microscope equipped with a DP71 camera. Select slides were also visualized using a Zeiss LSM 710 confocal microscope with Zen 2011 software.
Aiello N.M., Maddipati R., Norgard R.J., Balli D., Li J., Yuan S., Yamazoe T., Black T., Sahmoud A., Furth E.E., Bar-Sagi D, & Stanger B.Z. (2018). EMT subtype influences epithelial plasticity and mode of cell migration. Developmental cell, 45(6), 681-695.e4.
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