Anti-Claudin 3 antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 3 antibody (AB52231)

IHC image of ab52231 staining in normal human colon formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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

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Immunocytochemistry/ Immunofluorescence - Anti-Claudin 3 antibody (AB52231)

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.

• WB

AbReview29493****

Western blot - Anti-Claudin 3 antibody (AB52231)

Blocked with 0.1% Milk for 1 hour at 22°C

All lanes:

Western blot - Anti-Claudin 3 antibody (ab52231)

Lane 1:

Human urothelial cells (differentiated) at 15 µg

Lane 2:

Human urothelial cells (non-differentiated) at 15 µg

Secondary

All lanes:

IRDye® 800-conjugated Goat anti-rabbit IgG polyclonal

Predicted band size: 23 kDa

true

Exposure time: 2min

This image is courtesy of an anonymous Abreview

• WB

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Western blot - Anti-Claudin 3 antibody (AB52231)

All lanes:

Western blot - Anti-Claudin 3 antibody (ab52231) at 1/500 dilution

Lane 1:

Extracts from HuvEc cells, untreated.

Lane 2:

Extracts from HuvEc cells, treated with the immunizing peptide.

Predicted band size: 23 kDa

Observed band size: 23 kDa

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