Anti-Claudin 3 antibody

• Supplier: Abcam
• Catalog: ab52231
• Prices: $397.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ELISA WB ICC/IF ICC/IF IP IHC-P
• Species Reactivities: Human
• Antigen: Synthetic peptide derived from within amino acids 181 and 220 of Human Claudin-3
• Description: Rabbit Polyclonal
• Gene: CLDN3
• Conjugate: Unconjugated

Product images

All lanes : Anti-Claudin 3 antibody (ab52231) at 1/500 dilutionLane 1 : Extracts from HuvEc cells, untreated.Lane 2 : Extracts from HuvEc cells, treated with the immunizing peptide.

IHC image of ab52231 staining in normal human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52231, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ICC/IF image of ab52231 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52231, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

All lanes : Anti-Claudin 3 antibody (ab52231)Lane 1 : Human urothelial cells (differentiated)Lane 2 : Human urothelial cells (non-differentiated)Lysates/proteins at 15 µg per lane.SecondaryIRDye® 800-conjugated Goat anti-rabbit IgG polyclonaldeveloped using the ECL techniquePerformed under reducing conditions.

Claudin 3 was immunoprecipitated using 0.5mg MCF7 Whole cell lysate, 5µg of Rabbit polyclonal to Claudin 3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, MCF7 Whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52231.Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.Band: 18kDa; Claudin 3

Product References

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CD24 regulated gene expression and distribution of tight junction proteins is - CD24 regulated gene expression and distribution of tight junction proteins is

Ye P, Nadkarni MA, Simonian M, Hunter N. BMC Cell Biol. 2009 Jan 13;10:2.