Recombinant Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)
![Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)](https://www.abcam.com/ps/products/109/ab109110/Images/ab109110-296577-anti-eea1-antibody-epr4245-early-endosome-marker-western-blot.jpg "Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4245] to EEA1 - Early Endosome Marker
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human, African green monkey
Related conjugates and formulations
Overview
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Product name
Anti-EEA1 antibody [EPR4245] - Early Endosome Marker
See all EEA1 primary antibodies -
Description
Rabbit monoclonal [EPR4245] to EEA1 - Early Endosome Marker -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human, African green monkey -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: COS-1, NIH 3T3, C6, HeLa, Jurkat, Daudi, SH-SY5Y and JAR cell lysates. ICC/IF: JAR cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4245 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab185039)
- Alexa Fluor® 647 Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab196186)
- Alexa Fluor® 594 Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab206913)
- Anti-EEA1 antibody [EPR4245] - BSA and Azide free (ab239942)
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109110 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (1) |
1/500 - 1/1000.
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| WB | (2) |
1/10000 - 1/50000. Detects a band of approximately 170 kDa (predicted molecular weight: 162 kDa).
|
| Notes |
|---|
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ICC/IF
1/500 - 1/1000. |
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WB
1/10000 - 1/50000. Detects a band of approximately 170 kDa (predicted molecular weight: 162 kDa). |
Target
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Function
Binds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking. -
Sequence similarities
Contains 1 C2H2-type zinc finger.
Contains 1 FYVE-type zinc finger. -
Domain
The FYVE-type zinc finger domain mediates interactions with phosphatidylinositol 3-phosphate in membranes of early endosomes and penetrates bilayers. The FYVE domain insertion into PtdIns(3)P-enriched membranes is substantially increased in acidic conditions. -
Cellular localization
Cytoplasm. Early endosome membrane. - Information by UniProt
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Database links
- Entrez Gene: 8411 Human
- Entrez Gene: 216238 Mouse
- Entrez Gene: 314764 Rat
- Omim: 605070 Human
- SwissProt: Q15075 Human
- SwissProt: Q8BL66 Mouse
- Unigene: 567367 Human
- Unigene: 210035 Mouse
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Alternative names
- Early endosome antigen 1 antibody
- Early endosome antigen 1, 162kD antibody
- Early endosome associated protein antibody
see all
Images
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Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Early Endosome Marker knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: NIH3T3 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab109110 observed at 162 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab109110 was shown to recognize Early Endosome Marker in wild-type HAP1 cells as signal was lost at the expected MW in Early Endosome Marker knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Early Endosome Marker knockout samples were subjected to SDS-PAGE. Ab109110 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)](https://www.abcam.com/ps/products/109/ab109110/Images/ab109110-315314-anti-eea1-antibody-epr4245-early-endosome-marker-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)Immunocytochemistry/Immunofluorescence analysis of JAR (human placenta choriocarcinoma epithelial) cells labelling EEA1 with ab109110 at a dilution of 1/250. Cells were fixed with 4% paraformaldehye and permeabilized with 0.1% TritonX-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody at a dilution of 1/1000. Counterstained with DAPI and ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), at a dilution of 1/200.
Image shows cytoplasmic staining in JAR cell line.
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![Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)](https://www.abcam.com/ps/products/109/ab109110/Images/ab109110-447954-anti-eea1-antibody-epr4245-early-endosome-marker-western-blot-hela-edited-daudi-shsy5y-lysates.jpg)
Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)All lanes : Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EEA1 CRISPR/Cas9 edited HeLa cell lysate
Lane 3 : Daudi cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 162 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab109110 observed at 175 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109110 was shown to react with EEA1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab261822 (CRISPR/Cas9 edited cell lysate ab256897) lane below 175kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and EEA1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109110 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)](https://www.abcam.com/ps/products/109/ab109110/Images/ab109110-121675-anti-eea1-antibody-epr4245-early-endosome-marker-western-blot.jpg)
Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)All lanes : Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110) at 1/10000 dilution
Lane 1 : COS-1 cell lysate
Lane 2 : NIH 3T3 cell lysate
Lane 3 : C6 cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : Jurkat cell lysate
Lane 6 : JAR cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 162 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted? -
![Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)](https://www.abcam.com/ps/products/109/ab109110/Images/ab109110-4-benefits-of-recombinant-antibodies.png)
Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab109110 has been referenced in 13 publications.
- Sousa de Almeida M et al. Cellular Uptake of Silica and Gold Nanoparticles Induces Early Activation of Nuclear Receptor NR4A1. Nanomaterials (Basel) 12:N/A (2022). PubMed: 35215018
- Reyes-Alcaraz A et al. A NanoBiT assay to monitor membrane proteins trafficking for drug discovery and drug development. Commun Biol 5:212 (2022). PubMed: 35260793
- Wolf B et al. Therapeutic antibody glycosylation impacts antigen recognition and immunogenicity. Immunology 166:380-407 (2022). PubMed: 35416297
- Pan PY et al. Synaptojanin1 deficiency upregulates basal autophagosome formation in astrocytes. J Biol Chem 297:100873 (2021). PubMed: 34126070
- Hung C et al. SORL1 deficiency in human excitatory neurons causes APP-dependent defects in the endolysosome-autophagy network. Cell Rep 35:109259 (2021). PubMed: 34133918