Anti-Ezrin antibody [EP924Y] (ab75840)
![Anti-Ezrin antibody [EP924Y] (ab75840)](https://www.abcam.com/ps/products/75/ab75840/Images/ab75840-448926-anti-ezrin-antibody-ep924y-western-blot-hela-lysates.jpg "Anti-Ezrin antibody [EP924Y] (ab75840)")
Key features and details
- Rabbit monoclonal [EP924Y] to Ezrin
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
-
Product name
Anti-Ezrin antibody [EP924Y]
See all Ezrin primary antibodies -
Description
Rabbit monoclonal [EP924Y] to Ezrin -
Host species
Rabbit -
Specificity
ab75840 detects both phosphorylated and non phosphorylated Ezrin. -
Tested Applications & Species
See all applications and species dataApplication Species WB Human
-
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, HAP1, HCT116, 293, A431 and SH-SY5Y cell lysates.
-
General notes
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EP924Y -
Isotype
IgG -
Research areas
Images
-
Western blot - Anti-Ezrin antibody [EP924Y] (ab75840)All lanes : Anti-Ezrin antibody [EP924Y] (ab75840) 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 - ab75840 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab75840 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. ab75840 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.
-
![Western blot - Anti-Ezrin antibody [EP924Y] (ab75840)](https://www.abcam.com/ps/products/75/ab75840/Images/ab75840-296573-anti-ezrin-antibody-ep924y-western-blot.jpg)
Western blot - Anti-Ezrin antibody [EP924Y] (ab75840)All lanes : Anti-Ezrin antibody [EP924Y] (ab75840) at 1/1000 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 - ab75840 observed at 75 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab75840 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. Ab75840 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 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.
-
![Western blot - Anti-Ezrin antibody [EP924Y] (ab75840)](https://www.abcam.com/ps/products/75/ab75840/Images/ab75840-90963-anti-ezrin-antibody-ep924y-western-blot.gif)
Western blot - Anti-Ezrin antibody [EP924Y] (ab75840)All lanes : Anti-Ezrin antibody [EP924Y] (ab75840) at 1/10000 dilution
Lane 1 : 293 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : 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
Observed band size: 81 kDa why is the actual band size different from the predicted?
