PA1-063A

antibody from Invitrogen Antibodies

Targeting: EEA1 ZFYVE2

Provider product page for PA1-063A

Western blot

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [31]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [9]

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Product number: PA1-063A - Provider product page
Provider: Invitrogen Antibodies
Product name: EEA1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-063A detects early endosomal antigen 1 (EEA1) from mouse and human samples. Previous lots of this antibody have been used on hamster, canine, and rat samples. PA1-063A has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~160 kDa protein representing EEA1 from mouse brain extract. Previous lots of this antibody have been used in immunoprecipitation and immunofluorescence procedures. The PA1-063A immunogen is a synthetic peptide corresponding to residues T(1391) P S S K K P V R V C D A C F N D L Q G(1410) of human EEA1. The PA1-063A immunizing peptide (Cat. # PEP-079) is available for use in neutralization and control experiments.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

EEA1 protein structure - PA1-063A shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Cleavage of SNX2 disrupts interaction with Vps35. ( a ) HeLa cells were transfected for 16 h with plasmids encoding Cherry-fused full-length SNX2 (WT), C-terminal fragment (Delta84) or N-terminal fragment (1-84). Cherry-fused protein integrity was analyzed by immunoblotting using an RFP antibody (top). Colocalization with early endosomes was analyzed by confocal microscopy using SNX2, EEA1 early endosome marker and RFP antibodies (bottom). ( b ) HEK293T cells were transfected for 48 h with the indicated combination of empty (Myc), Myc-tagged Vps35, SNX2 WT and SNX2Delta84. Samples were immunoprecipitated (IP) using a Myc antibody and proteins were analyzed by immunoblotting (IB) with the indicated antibodies. ( c ) HeLa cells were transfected two times at a 24 h interval with the indicated combination of control (CTL), SNX1 and SNX2 small interfering RNAs (siRNAs), and proteins were analyzed after 72 h by immunoblotting using the indicated antibodies. Actin was used as a loading control. ( d ) Mixed populations of cells transfected as in ( c ) were analyzed for ci-MPR, SNX2 and Vps26 localization by confocal microscopy. Arrows point to SNX1/SNX2-depleted cells. Scale bars in ( a ) and ( d ) represent 10 mu m.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Subcellular localization of TLR8 in human monocytes and HeLa transfectants. HeLa cells transiently expressing human TLR8 ( A ) and human monocytes ( B ) were incubated with anti-FLAG mAb ( A ) or anti-TLR8 mAb ( B ) followed by an Alexa Fluor 488-conjugated secondary Ab. Organelles were stained with an anti-EEA1 pAb, anti-MPR pAb, anti-p115 pAb, anti-calreticulin pAb, or anti-calnexin pAb followed by an Alexa Fluor 568-conjugated secondary Ab. Representative confocal images are shown. Green, TLR8; red, organelle markers; blue, nuclei stained with DAPI. Scale bar: ( A ) 10 um; ( B ) 5 um.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Defining the TLR domain responsible for localization. A , Schematic diagram of the tail-deletion constructs of hTLR8. B , Expression of the TLR tail-deletion constructs in HEK293FT cells. Wild-type and mutant proteins transiently expressed in HEK293FT cells were immunoprecipitated with anti-FLAG mAb, resolved using SDS-PAGE, and detected using immunoblotting with anti-FLAG mAb. Molecular weight markers are shown on the right. C , Immunofluorescence images of the TLR8 deletion constructs in transfected HeLa cells. The upper and middle panels show cells stained with delTIR and delCYT together with phalloidin, which labels the plasma membrane. The lower two panels show cells stained with GPI-TLR8 together with EEA1 or calreticulin. Green, TLR8 mutants; red, organelles; blue, nuclei stained with DAPI; bar, 10 um.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 5 Endosomal localization of Cytc-Tf conjugate in A549 cells using confocal imaging. (A) A549 cells were treated with the Cyt c-Tf conjugate for 12 h and endosome localization was probed with an endosomal marker antibody (primary antibody against EEA1 and a secondary antibody conjugated with Alexafluor-555) and Cyt c was localized with an anti-Cyt c FITC labeled antibody. The upper panel shows untreated cells and the lower panel the cells treated with Cyt c-Tf. (B) Merged image of A549 cells treated with conjugate enlarged to show nonover lapping green (Cyt c) and red (endosomes) signal.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Characterization of the compartments involved in the trafficking of IgG. ( A ) Representative confocal microscopy images showing the distribution of early endosomes (EEA1, cyan), recycling endosomes (Rab11, green), and lysosomes (LAMP1, orange) in diploid (NDS) and trisomic (DS) fibroblasts after incubation with IgG for 60 min at 37 degC. Scale bar: 10 um. ( B ) Number and size of EEA1 + endosomes (upper panel), Rab11 + recycling endosomes (upper middle panel), and lysosomes (LAMP1, Lysotracker) (lower middle and lower panels) in NDS (NDS-1, -2, and -3) and DS (DS-1, -2, and -3) cells. Each bar represents the mean +- SD. Each point represents measurements in individual cells (NDS, 72-296 cells/marker; DS, 82-250 cells cells/marker). **P < 0.01, ***P < 0.001, ns = not significant, Student's t test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Analyses of representative samples of SH-SY5Y cells expression of DAPI, Vimentin and EEA1 were shown. Immunochemical stains for DAPI (blue), vimentin (green) and EEA1 (red) for SH-SY5Y cells cultured with aluminum ion and NAP for 24 h. (Scale bars, 10 mum, Confocal microscope, 400X).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 8 C . burnetii T4BSS alters latex bead phagosome maturation. Mock and C . burnetii -infected murine alveolar macrophages (2 dpi) were pulsed with 1 mum red fluorescent latex beads for 15 min, followed by fixation at 0 min post-pulse and every 15 min for 60 min. Cells were then immunostained for C . burnetii , plasma membrane, and Rab5 or Rab7 and internalized beads scored for co-localization with Rab5 or Rab7. The percent of Rab5 or Rab7 positive bead phagosomes over total number of internalized beads was plotted against time. In WT-infected cells, Rab5 was lost from bead phagosomes significantly faster than mock and DeltadotA -infected cells (A), whereas Rab7 localization to bead phagosomes was significantly slower in WT-infected cells (B). Further, bead phagosomes in mock, WT, and DeltadotA -infected cells were immunostained for EEA1, LAMP1, and Rab11 at 0 min and 60 min post-pulse. At 60 min post pulse, significantly more bead phagosomes retained EEA1, whereas WT-infected cells retained significantly more EEA1 compared to mock and DeltadotA (C), significantly fewer of those were positive for LAMP1 at 60 min post-pulse (D). There was no difference in Rab11 profile of bead phagosomes in between mock, WT, and DeltadotA -infected cells. Data shown as mean+-SEM of at least 30 bead phagosomes per condition in each of three independent experiments as analyzed by multiple t-test (A, B) or one-way ANOVA (C-E); **, P