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Rabbit anti eea1

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Rabbit anti-EEA1 is a primary antibody that recognizes the Early Endosome Antigen 1 (EEA1) protein. EEA1 is a marker for early endosomes and is involved in endosome fusion and trafficking processes. This antibody can be used for immunofluorescence, immunohistochemistry, and Western blot applications to detect and localize the EEA1 protein.

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Lab products found in correlation

Rabbit anti vwf, by Agilent Technologies (2 mentions) Rabbit anti lamp1, by Cell Signaling Technology (2 mentions) Mouse anti eea1, by BD (2 mentions) Mouse pi 3 4 p2 igg, by Echelon Biosciences (2 mentions) Mouse pi 4 5 p2 igm, by Echelon Biosciences (2 mentions) Mouse pi 4 p igm, by Echelon Biosciences (2 mentions) Mouse pi 3 p igg, by Echelon Biosciences (2 mentions) Mouse anti gfp, by Takara Bio (2 mentions) Mouse anti gapdh, by Merck Group (2 mentions) Sheep anti vwf, by Abcam (1 mentions) Anti human cd31, by Agilent Technologies (1 mentions) Mouse anti lamp1, by Abcam (1 mentions) Rabbit anti rab9, by Cell Signaling Technology (1 mentions) F actin, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit alexa 488, by Thermo Fisher Scientific (1 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Dynasore, by Merck Group (1 mentions) Texas red dextran, by Thermo Fisher Scientific (1 mentions) Anti rab11, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Anti-EEA1 (37 citations) Rabbit anti-EEA1 antibody (5 citations) Rabbit anti-EEA1 (C45B10) (4 citations)

16 protocols using rabbit anti eea1

1

Immunohistochemical Profiling of Cells

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Antibodies used include: sheep anti-FVIII (Affinity Biologicals), rabbit anti-VWF (Dako), sheep anti-VWF (Abcam, Cambridge, UK), rabbit anti-EEA1 (Cell Signalling Technology, Beverly, MA, USA), rabbit anti-SCARA5 (HPA024661, Sigma Aldrich), mouse anti-SCARA5 (αh-SR5.2), anti-human CD31 (Dako), anti-human CD68 (Santa Cruz Biotechnology, Dallas, USA), anti-human synaptopodin (R&D, Minneapolis, USA), mouse anti-human CD34 (QBEnd-10, ThermoFisher Scientific, Waltham, USA) and mouse anti-human CD8α (4B11, ThermoFisher Scientific).
Swystun L.L., Ogiwara K., Lai J.D., Ojala J.R., Rawley O., Lassalle F., Notley C., Rengby O., Michels A., Nesbitt K., Tryggvason K, & Lillicrap D. (2019). The scavenger receptor SCARA5 is an endocytic receptor for von Willebrand factor expressed by littoral cells in the human spleen. Journal of thrombosis and haemostasis : JTH, 17(8), 1384-1396.
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2

Immunohistochemical Antibody Staining

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Antibodies used included: sheep anti-F8 (Affinity Biologicals), rabbit anti-VWF (DAKO), mouse anti-CLEC4M (R&D), rabbit anti-CLEC4M (Novus, Littleton, USA), rabbit anti-EEA1 (Cell Signalling Technology, Beverly, MA, USA), rabbit anti-Rab9 (Cell Signaling Technology), mouse anti-LAMP1 (Abcam), and rat anti-murine CD31 (Dianova, Hamburg, Germany).
Swystun L., Notley C., Georgescu I., Lai J., Nesbitt K., James P, & Lillicrap D. (2019). The endothelial lectin clearance receptor CLEC4M binds and internalizes Factor VIII in a VWF-dependent and -independent manner. Journal of thrombosis and haemostasis : JTH, 17(4), 681-694.
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3

Myelin Internalization and Endosomal Trafficking in Confluent BECs

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Confluent BECs were seeded in 8-well Ibidi slides (Ibidi, GmbH, Munchen, Germany) and incubated with 10 µg/ml Atto 633 labeled myelin for 24 hr. Subsequently, cells were extensively washed with PBS and fixated with 4% formaldehyde. Non-specific binding was blocked with 5% goat serum in PBS/BSA containing 0.3% Triton-X100. Cells were labeled with rabbit anti-EEA1 (Cell Signaling) or rabbit anti-LAMP1 (Cell Signaling). Antibodies were visualized after 1 hr incubation with goat anti-rabbit Alexa488 (Molecular Probes). Finally, sections were stained with Hoechst (molecular Probes, Invitrogen) to visualize cellular nuclei and with phalloidin rhodamine to visualize F-actin (Molecular Probes, Invitrogen). Sections were mounted with mounting medium. Co-localization was analyzed using a Confocal Laser Scanning Microscope (Leica DMI 6000, SP8, Leica, Mannheim, Germany); images were acquired using LCS software (version 2.61, Leica).
Lopes Pinheiro M.A., Kamermans A., Garcia-Vallejo J.J., van het Hof B., Wierts L., O'Toole T., Boeve D., Verstege M., van der Pol S.M., van Kooyk Y., de Vries H.E, & Unger W.W. (2016). Internalization and presentation of myelin antigens by the brain endothelium guides antigen-specific T cell migration. eLife, 5, e13149.
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4

Endocytic Trafficking Mechanisms Study

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Dynasore was obtained from Sigma-Aldrich. AlexaFluor 488-albumin from bovine serum conjugate, TexasRed-dextran (70,000 MW) lysine fixable, and AlexaFluor594-transferrin were from ThermoFisher Scientific. AlexaFluor594-phalloidin was obtained from ThermoFisher Scientific. The following antibodies and reagents were used: rabbit anti-prosurfactant protein C (Abcam, Cambridge, UK), rat anti-RAGE (R&D Systems, Minneapolis, MN, USA), rabbit anti-EEA1, anti-Rab11, and anti-clathrin from Cell Signaling (Cell Signaling, Danvers, MA, USA).
Kryvenko V., Alberro-Brage A., Fysikopoulos A., Wessendorf M., Tello K., Morty R.E., Herold S., Seeger W., Samakovlis C, & Vadász I. (2023). Clathrin-Mediated Albumin Clearance in Alveolar Epithelial Cells of Murine Precision-Cut Lung Slices. International Journal of Molecular Sciences, 24(3), 2644.
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5

Immunofluorescent Analysis of BM-DC Endosomes

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BM-DCs were grown on Bio-coat coverslips (BD biosciences), fixed in 4% paraformaldehyde, permeabilized with 0.1% saponin and blocked with 5% goat serum. Samples were incubated with anti-CD11b-V450 (BD Biosciences, dilution 1:50), anti-CD14-biotin (ebioscience; dilution 1:50) and rabbit anti-EEA1 (Cell Signaling Technology; dilution 1:100). Secondary antibodies Alexa Fluor 555-conjugated Streptavidin and Alexa Fluor 488-conjugated goat-anti-rabbit IgG were both from Molecular Probes and were added at a dilution of 1:100. Samples were then analysed with a Leica SP5 confocal microscope. Colocalization spectra of each channel in the region of interest were analysed using the Lecia LAS AF software.
Ling G.S., Bennett J., Woollard K.J., Szajna M., Fossati-Jimack L., Taylor P.R., Scott D., Franzoso G., Cook H.T, & Botto M. (2014). Integrin CD11b positively regulates TLR4-induced signalling pathways in dendritic cells but not in macrophages. Nature Communications, 5, 3039.
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6

Immunofluorescence Staining Reagents and Plasmids

Cited 1 time
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Rabbit anti-Hrs was raised using GST–Hrs as an immunogen and was generated by Eurogentec, Southampton, UK. The antibody was affinity-purified against GST–Hrs and adsorbed against GST. Sheep anti-VPS28 was as described previously (Bishop et al., 2002 (link)). Mouse anti-Hrs was from Enzo. Rabbit anti-EEA1 was from Cell Signaling Technology. Mouse anti-EEA1 was from BD Biosciences. Mouse anti-EGFR was Mab 108, purified from supernatants of the hybridoma clone HB-9764 (ATCC; Washington DC), grown in RPMI medium and 10% FCS under 5% CO2, and antibody affinity-purified using Protein-G–Sepharose. Rabbit anti-APPL1 and anti-CHMP4B were from Proteintech Europe (Manchester, UK). Rabbit anti-clathrin was from Abcam (Cambridge, UK). Fluorescent EGF was from Invitrogen (Paisley, Scotland) and secondary antibodies were from Jackson ImmunoResearch (West Grove, PA). Human SNX15 in pEGFP-C1 (Clontech) was a gift from Chris Danson and Pete Cullen, University of Bristol.
Flores-Rodriguez N., Kenwright D.A., Chung P.H., Harrison A.W., Stefani F., Waigh T.A., Allan V.J, & Woodman P.G. (2015). ESCRT-0 marks an APPL1-independent transit route for EGFR between the cell surface and the EEA1-positive early endosome. Journal of Cell Science, 128(4), 755-767.
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7

Antibody Profiling of EHD and EEA1

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The following antibodies were used: Rabbit anti-EHD1 (Abcam, ab109311), Rabbit anti-EHD4 [32 (link)], Rabbit anti-EEA1 (Cell Signaling, #3288), Rabbit anti-HA (SAB, #T501), Mouse anti-GFP (Roche, 11814460001), Mouse anti-LRP1 (Novus, NB100-64808), Donkey anti-mouse-HRP (Jackson, 715-035-151), Mouse anti-rabbit IgG light chain-HRP (Jackson, 211-032-171), Alexa-fluor 568-conjugated goat anti-rabbit (Molecular Probes, A11036).
Jones T., Naslavsky N, & Caplan S. (2020). Eps15 Homology Domain Protein 4 (EHD4) is required for Eps15 Homology Domain Protein 1 (EHD1)-mediated endosomal recruitment and fission. PLoS ONE, 15(9), e0239657.
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8

Antibody Reagents for Cellular Signaling

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rabbit anti‐RPTPγ (rb‐RPTPγ‐P4: gift from C. Sorio, Department of Pathology, University of Verona, Verona, Italy, (Sorio et al, 1995 (link))), mouse anti‐TCPTP (MAB1930, R&D Systems, Minneapolis, MN), mouse anti‐p22phox (ab80896, Abcam, Cambridge, UK), anti‐cysteine sulfenic acid 2‐thiodimedone (ABS30, Merck, Darmstadt, Germany), living colors mouse anti‐GFP (632381, Clontech, Mountain View, CA), goat anti‐GFP (ab5450, Abcam), rabbit anti‐phospho EGFR Y1068 (3777, Cell Signaling Technology, Danvers, MA), mouse anti‐phospho EGFR Y1068 (2236, Cell Signaling Technology) rabbit anti‐EGFR (4267, Cell Signaling Technology), goat anti‐EGFR (AF231, R&D Systems), rabbit anti‐phospho‐ERK‐1/2 Thr/Tyr 202/204 (9101, Cell Signaling Technology), mouse anti‐ERK1/2 (ab366991, Abcam), rabbit anti‐phospho‐Akt Ser473 (9271, Cell Signaling Technology), mouse anti‐Akt (pan) (2920, Cell Signaling Technology), rabbit anti‐EEA1 (3288, Cell Signaling Technology), rabbit anti‐Rab7 (9367, Cell Signaling Technology), rabbit anti‐Rab11a (2413, Cell Signaling Technology), rabbit phospho‐Rb Ser807/811 (8516, Cell Signaling Technology), mouse anti‐GAPDH (CB1001, Merck), mouse anti‐α‐Tubulin (T6074, Merck), mouse anti‐Na+/K+ ATPase‐α3‐subunit (BML‐SA247‐0100, Enzo Life Sciences, NY).
Joshi M.S., Stanoev A., Huebinger J., Soetje B., Zorina V., Roßmannek L., Michel K., Müller S.A, & Bastiaens P.I. (2023). The EGFR phosphatase RPTPγ is a redox‐regulated suppressor of promigratory signaling. The EMBO Journal, 42(10), e111806.
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9

Antibody Panel for Protein Analysis

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The following antibodies were used: mouse anti-EEA1, rabbit anti-Crk (BD Biosciences), rabbit anti-ElgB, rabbit anti-GAPDH, rabbit anti-Cul5, mouse anti-Cul5, mouse anti-IRS4, mouse anti-Myc (9E10) and mouse anti-tubulin (Santa Cruz Biotechnology, Inc.), rabbit anti-beta catenin, mouse anti-Flag (M2) (Sigma-Aldrich), mouse anti-EphA2 (8B6), rabbit anti-EphA2 (D4A2), rabbit anti-EphA2 (pY588), rabbit anti-EphA2 (pY772), rabbit anti-EEA1, rabbit anti-Ezrin (Cell Signaling Technology); mouse anti-T7 (EMD Millipore), mouse anti-HA (HA.11) (BioLegend), rabbit anti-HA (Bethyl) and goat anti-human IgG Fc (Jackson ImmunoResearch Inc.). 4G10 was made in-house from the 4G10 hybridoma.
Pilling C, & Cooper J.A. (2017). SOCS2 Binds to and Regulates EphA2 through Multiple Mechanisms. Scientific Reports, 7, 10838.
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10

Antibody Immunostaining Protocol for Organelles

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The primary antibodies used in this study were: rabbit anti-Rab7 (for IHC: ab 137029; Abcam, Cambridge, UK), chicken anti-Rab7 (for WB), rabbit anti-LC3 (for WB: PM036; MBL, Nagoya, Japan) (for IHC: #3868; Cell Signaling Technology), guinea pig anti-p62 (PM066; MBL), rat anti-N-terminal of lysosomal-associated membrane protein (LAMP)-1 (553792; BD Biosciences, San Jose, CA, USA) and rabbit anti-C-terminal of LAMP-1 (ab24170; Abcam), mouse anti-cathepsin B (AF965; R&D Systems, Minneapolis, MN, USA), rabbit anti-EEA1 (#2411; Cell Signaling Technology), mouse anti-CI-MPR (NB300–514; Novus, Littleton, CO, USA) and goat anti-actin (sc-1616; Santa Cruz Biotechnology, Dallas, TX, USA) immunoglobulins (IgGs). The secondary antibodies used in this study were: horseradish peroxidase-conjugated goat anti-guinea pig (Cosmo Bio, Tokyo, Japan), donkey anti-rabbit, anti-chicken and anti-mouse IgGs, and FITC- or Cy3-conjugated donkey anti-rabbit, anti-mouse and anti-rat IgGs (Jackson ImmunoResearch, West Grove, PA, USA).
Takahashi K., Mashima H., Miura K., Maeda D., Goto A., Goto T., Sun-Wada G.H., Wada Y, & Ohnishi H. (2017). Disruption of Small GTPase Rab7 Exacerbates the Severity of Acute Pancreatitis in Experimental Mouse Models. Scientific Reports, 7, 2817.
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