Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 09, 2021

Anti-EpCAM antibody [EPR20533-266] (ab213501)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20533-266] to EpCAM
Suitable for: WB, ICC/IF, IP, IHC-P
Reacts with: Mouse, Rat

Product name

Anti-EpCAM antibody [EPR20533-266]
See all EpCAM primary antibodies
Description

Rabbit monoclonal [EPR20533-266] to EpCAM
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Mouse
IHC-P	Mouse Rat
IP	Mouse
WB	Mouse Rat

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse small intestine, pancreas, colon and kidney lysates; Rat colon and kidney lysates; 4T1 whole cell lysate. IHC-P: Mouse colon and lung tissues; Rat colon tissue. ICC/IF: 4T1 cells. IP: 4T1 whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20533-266
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20533-266] (ab213501)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling EpCAM with ab213501 at 1/2000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat colon is observed (PMID: 15637741). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [EPR20533-266] (ab213501)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 4T1 (mouse mammary gland carcinoma cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling EpCAM with ab213501 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on 4T1 cells.

Negative control: NIH/3T3 (PMID:23264216).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20533-266] (ab213501)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling EpCAM with ab213501 at 1/2000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse lung is observed (PMID: 15637741). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20533-266] (ab213501)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling EpCAM with ab213501 at 1/2000 dilution, followed by Goat anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse colon is observed (PMID: 15637741). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-EpCAM antibody [EPR20533-266] (ab213501)

Lanes 1-3 : Anti-EpCAM antibody [EPR20533-266] (ab213501) at 1/1000 dilution
Lanes 4-6 : Anti-EpCAM antibody [EPR20533-266] (ab213501) at 1/5000 dilution

Lane 1 : Mouse small intestine lysate
Lane 2 : Mouse pancreas lysate
Lane 3 : Rat colon lysate
Lane 4 : Mouse colon lysate
Lane 5 : Mouse kidney lysate
Lane 6 : Rat kidney lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 35 kDa
Observed band size: 39 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-3: 3 minutes; Lane 4/5: 5 seconds; Lane 6: 30 seconds.

The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).
Western blot - Anti-EpCAM antibody [EPR20533-266] (ab213501)

All lanes : Anti-EpCAM antibody [EPR20533-266] (ab213501) at 1/5000 dilution

Lane 1 : 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 35 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).

Negative control:

NIH/3T3 (PMID:23264216).
Immunoprecipitation - Anti-EpCAM antibody [EPR20533-266] (ab213501)

EpCAM was immunoprecipitated from 0.35 mg of 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate with ab213501 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab213501 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab13136), was used as secondary antibody at 1/10000 dilution.

Lane 1: 4T1 whole cell lysate 10 µg (Input).

Lane 2: ab213501 IP in 4T1 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213501 in 4T1 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Anti-EpCAM antibody [EPR20533-266] (ab213501)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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