ET1609-70

Pan-Cadherin Recombinant Rabbit Monoclonal Antibody [ST54-01]

cat.: ET1609-70

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, IHC-Fr
Clonality:	Monoclonal
Clone number:	ST54-01

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 100 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within C-terminal human CDH2.
Positive control:	Human heart tissue lysate, mouse heart tissue lysate, rat heart tissue lysate, HeLa, Caco-2, mouse liver tissue, mouse kidney tissue, mouse heart tissue.
Subcellular location:	Cell membrane, Cell junction, Endosome, Golgi apparatus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP IHC-Fr	1:1,000 1:50-1:200 1:200 1:50-1:1,000 1:50-1:100 Use at an assay dependent concentration. 1:200
Uniprot #:	SwissProt: P19022 Human \| P12830 Human \| P22223 Human \| P33151 Human \| P55283 Human \| P39038 Mouse
Alternative names:	Cadherin CDH3 CDHP 7B4 antigen Cadherin-1 Cadherin-2 Cadherin-3 Cadherin-4 Cadherin-5 CAM 120/80 CD144 CD324 CD325 Cdh4 CDH5 CDHE CDHN E-cadherin Epithelial cadherin N-cadherin NCAD Neural cadherin P-cadherin Placental cadherin UVO Uvomorulin Vascular endothelial cadherin

Images

Fig1: Western blot analysis of Pan-Cadherin on different lysates with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/1,000 dilution.

Lane 1: Human heart tissue lysate
Lane 2: Mouse heart tissue lysate
Lane 3: Rat heart tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 100 kDa
Observed band size: 130 kDa

Exposure time: 1 minute;

6% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-70) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/300,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HeLa cells labeling Pan-Cadherin with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunocytochemistry analysis of Caco-2 cells labeling Pan-Cadherin with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-70) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-70) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-70) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunofluorescence analysis of paraffin-embedded mouse liver tissue labeling Pan-Cadherin with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-70, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig8: Immunofluorescence analysis of frozen mouse liver tissue with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-70, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig9: Flow cytometric analysis of Pan-Cadherin was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-70, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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