Anti-Ezrin antibody [EPR803] (ab133297)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR803] to Ezrin
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Ezrin antibody [EPR803]
See all Ezrin primary antibodies -
Description
Rabbit monoclonal [EPR803] to Ezrin -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Synthetic peptide within Human Ezrin aa 350-450. The exact sequence is proprietary.
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Positive control
- WB: HAP1, HeLa, HCT116, 293T, A549 and SH-SY5Y cell lysates. IHC-P: Human kidney tissue and Human transitional cell carcinoma tissue.
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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 -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR803 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EZR knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab133297 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133297 was shown to react with Ezrin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264912 (knockout cell lysate ab256908) was used. Wild-type HeLa and EZR knockout 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. ab133297 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Ezrin with ab133297 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Ezrin knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HCT116 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 69 kDaLanes 1 - 4: Merged signal (red and green). Green - ab133297 observed at 75 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133297 was shown to specifically react with Ezrin in wild-type HAP1 cells as signal was lost in Ezrin knockout cells. Wild-type and Ezrin knockout samples were subjected to SDS-PAGE. Ab133297 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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Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma tissue labelling Ezrin with ab133297 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : A549 cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-rabbit HRP at 1/2000 dilution
Predicted band size: 69 kDa
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