Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Claudin 3 antibody [EPR19971] (ab214487)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19971] to Claudin 3
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
Reacts with: Human

Product name

Anti-Claudin 3 antibody [EPR19971]
See all Claudin 3 primary antibodies
Description

Rabbit monoclonal [EPR19971] to Claudin 3
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: SW480, MCF7 and Caco-2 whole cell lysates; human breast cancer, pancreas, ovary cancer and fetal kidney lysates. IHC-P: Human pancreas, prostate and breast cancer tissues. ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells. IP: MCF7 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: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19971
Isotype

IgG
Research areas

Western blot - Anti-Claudin 3 antibody [EPR19971] (ab214487)

All lanes : Anti-Claudin 3 antibody [EPR19971] (ab214487) at 1/1000 dilution

Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 23 kDa
Observed band size: 20 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

Doublet bands observed correspond to full length and processed forms of the claudin-3 protein. This is consistent with what has been described in the literature (PMID: 26170992).
Western blot - Anti-Claudin 3 antibody [EPR19971] (ab214487)

All lanes : Anti-Claudin 3 antibody [EPR19971] (ab214487) at 1/1000 dilution

Lane 1 : Human breast cancer lysate
Lane 2 : Human pancreas lysate
Lane 3 : Human ovary cancer lysate
Lane 4 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/5000 dilution

Predicted band size: 23 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 1 minute; Lane 3: 3 seconds; Lane 4: 3 minutes.

Doublet bands observed correspond to full length and processed forms of the claudin-3 protein. This is consistent with what has been described in the literature (PMID: 26170992).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 3 antibody [EPR19971] (ab214487)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the pancreatic acinar cells of human pancreas is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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-Claudin 3 antibody [EPR19971] (ab214487)

Immunohistochemical analysis of paraffin-embedded human prostate tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the epithelium of human benign prostatic hyperplasia is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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-Claudin 3 antibody [EPR19971] (ab214487)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Claudin 3 with ab214487 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on the tumor cells of human breast cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Claudin 3 antibody [EPR19971] (ab214487)

Immunofluorescent analysis of 100% methanol-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Claudin 3 with ab214487 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 membrane staining on MCF7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-Claudin 3 antibody [EPR19971] (ab214487)

Flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Claudin 3 with ab214487 at 1/50 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Claudin 3 antibody [EPR19971] (ab214487)

Claudin 3 was immunoprecipitated from 0.35 mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab214487 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214487 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1: MCF7 whole cell lysate 10µg (Input).

Lane 2: ab214487 IP in MCF7 whole cell lysate.

Lane 3:Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214487 in MCF7 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.
Anti-Claudin 3 antibody [EPR19971] (ab214487)

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