Cadherin-Catenin Antibody Sampler Kit
cat.: HAK21157
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB, IHC-P
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Uniprot #: SwissProt: P35221 Human | P26231 Mouse | P35222 Human | Q02248 Mouse | Q9WU82 Rat | O60716 Human | Q9UQB3 Human | O35927 Mouse | O35116 Rat | P12830 Human | P09803 Mouse | Q9R0T4 Rat | P14923 Human | Q02257 Mouse | Q6P0K8 Rat | P19022 Human | P15116 Mouse | Q9Z1Y3 Rat | P19022 Human | P12830 Human | P22223 Human | P33151 Human | P55283 Human | P39038 Mouse
Entrez Gene: 307505 Rat
HAK21157 - Page 2
Images
HAK21157_1.jpg Fig1: Western blot analysis of N Cadherin on different lysates with Rabbit anti-N Cadherin antibody (ET1607-37) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: 293T cell lysate
Lane 2: A549 cell lysate
Lane 3: HeLa cell lysate
Lane 4: A-172 cell lysate
Lane 5: MCF7 cell lysate (negative)
Lane 6: C2C12 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 100 kDa
Observed band size: 140-150 kDa

Exposure time: 2 minutes 6 seconds; ECL: K1801;
4-20% 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 (ET1607-37) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157_2.jpg Fig2: Western blot analysis of N Cadherin on different lysates with Rabbit anti-N Cadherin antibody (ET1607-37) at 1/5,000 dilution.

Lane 1: 293T-si NT cell lysate (10 µg/Lane)
Lane 2: 293T-si N Cadherin cell lysate (10 µg/Lane)

Predicted band size: 100 kDa
Observed band size: 150 kDa

Exposure time: 1 minute 46 seconds; ECL: K1801;
4-20% 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 (ET1607-37) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157 - Page 3
HAK21157_3.jpg Fig3: Western blot analysis of Delta-catenin on different lysates with Rabbit anti-Delta-catenin antibody (HA722438) at 1/1,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse hippocampus tissue lysate
Lane 3: Mouse heart tissue lysate (negative)
Lane 4: Mouse kidney tissue lysate (negative)
Lane 5: Mouse cerebellum tissue lysate
Lane 6: Rat brain tissue lysate
Lane 7: Rat heart tissue lysate (negative)
Lane 8: Rat kidney tissue lysate (negative)
Lane 9: Rat spleen tissue lysate (negative)
Lane 10: Rat hippocampus tissue lysate
Lane 11: Rat cerebellum tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 135 kDa
Observed band size: 135 kDa

Exposure time: 1 minute 2 seconds; ECL: K1801;

4-20% 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 (HA722438) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157_4.jpg Fig4: 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.
HAK21157 - Page 4
HAK21157_5.jpg Fig5: Western blot analysis of E-Cadherin on different lysates with Rabbit anti-E-Cadherin antibody (HA723564) at 1/5,000 dilution.

Lane 1: 4T1 cell lysate (20 µg/Lane)
Lane 2: C2C12 cell lysate (negative) (20 µg/Lane)
Lane 3: Mouse small intestine tissue lysate (30 µg/Lane)
Lane 4: Mouse colon tissue lysate (30 µg/Lane)
Lane 5: Mouse pancreas tissue lysate (30 µg/Lane)
Lane 6: Rat pancreas tissue lysate (30 µg/Lane)
Lane 7: Rat lung tissue lysate (30 µg/Lane)
Lane 8: Rat colon tissue lysate (30 µg/Lane)
Lane 9: MCF7 cell lysate (20 µg/Lane)
Lane 10: MDA-MB-231 cell lysate (negative) (20 µg/Lane)
Lane 11: HT-29 cell lysate (20 µg/Lane)
Lane 12: Caco-2 cell lysate (20 µg/Lane)

Predicted band size: 98 kDa
Observed band size: 75-130 kDa

Exposure time: Lane 1-5: 10 seconds; Lane 3: 1 minute 16 seconds; ECL: K1801;

4-20% 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 (HA723564) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-N Cadherin antibody (ET1607-37) at 1/40,000 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 (ET1607-37) at 1/40,000 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.
HAK21157 - Page 5
HAK21157_7.jpg Fig7: Western blot analysis of Pan-Cadherin on different lysates with Rabbit anti-Pan-Cadherin antibody (ET1609-70) at 1/2,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: Mouse embryo tissue lysate (40 µg/Lane)
Lane 4: Mouse placenta tissue lysate (40 µg/Lane)
Lane 5: Rat embryo tissue lysate (40 µg/Lane)

Predicted band size: 100 kDa
Observed band size: 120 kDa

Exposure time: 4 seconds; ECL: K1801;

4-20% 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 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157_8.jpg Fig8: Fluorescence multiplex immunohistochemical analysis of mouse liver (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Tangerine), anti-αSMA (ET1607-53, Yellow), anti-SOX9 (ET1611-56, Green), anti-Albumin (ET1702-55, Cyan) anti-GS (EM1902-39, Magenta) and anti-CK19 (ET1601-6, Orange) on mouse liver. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in six rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1607-53 (1/3,000 dilution), ET1611-56 (1/1,500 dilution), ET1702-55 (1/3,000 dilution), EM1902-39 (1/2,000 dilution) and ET1601-6 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HAK21157 - Page 6
HAK21157_9.jpg Fig9: Fluorescence multiplex immunohistochemical analysis of mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Beta Catenin (ET1601-5, Violet), anti-Glucagon (ET1702-20, Green) and anti-Insulin (ET1601-12, White) on pancreas. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1702-20 (1/6,000 dilution) and ET1601-12 (1/8,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
HAK21157_10.jpg Fig10: Fluorescence multiplex immunohistochemical analysis of mouse liver (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Th (ET1611-12, Green), anti-HNF4a (HA721006, Magenta), anti-CK19 (ET1601-6, Cyan), anti-α-sma (ET1607-53, Red) and anti-β-catenin (ET1601-5, Yellow) on liver. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1611-12 (1/1,000 dilution), HA721006 (1/2,000 dilution), ET1601-6 (1/3,000 dilution), ET1607-53 (1/10,000 dilution) and ET1601-5 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HAK21157 - Page 7
HAK21157_11.jpg Fig11: Immunofluorescence analysis of paraffin-embedded mouse colon tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 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 (ET1601-5, green) at 1/100 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).
HAK21157_12.jpg Fig12: Immunocytochemistry analysis of A431 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) 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-Beta Catenin antibody (ET1601-5) 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.
HAK21157 - Page 8
HAK21157_13.jpg Fig13: Immunocytochemistry analysis of C6 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) 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-Beta Catenin antibody (ET1601-5) 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.
HAK21157_14.jpg Fig14: Immunofluorescence analysis of frozen mouse colon tissue with Rabbit anti-Beta Catenin antibody (ET1601-5) 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 (ET1601-5, 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).
HAK21157_15.jpg Fig15: Beta Catenin was immunoprecipitated from 0.2 mg rat brain tissue lysate with ET1601-5 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1601-5 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Rat brain tissue lysate (input)
Lane 2: ET1601-5 IP in rat brain tissue lysate
Lane 3: Rabbit IgG instead of ET1601-5 in rat brain tissue lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 2 seconds; ECL: K1801
HAK21157 - Page 9
HAK21157_16.jpg Fig16: Flow cytometric analysis of A431 cells labeling Beta Catenin.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-5, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HAK21157_17.jpg Fig17: Application: IHC-Fr

Species: Mouse

Site: colon

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: Not required
HAK21157_18.jpg Fig18: Application: IHC-Fr

Species: Rat

Site: colon

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: Not required
HAK21157 - Page 10
HAK21157_19.jpg Fig19: Immunocytochemistry analysis of MCF7 cells labeling E-Cadherin with Rabbit anti-E-Cadherin antibody (HA723564) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-E-Cadherin antibody (HA723564) 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 (HA601187, 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.
HAK21157_20.jpg Fig20: Flow cytometric analysis of C2C12 (left, negative) and 4T1 (right, positive) cells labeling E-Cadherin.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA723564, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HAK21157 - Page 11
HAK21157_21.jpg Fig21: Western blot analysis of E-Cadherin on different lysates with Rabbit anti-E-Cadherin antibody (HA723564) at 1/5,000 dilution.

Lane 1: His-tagged Mouse E-cadherin recombinant protein
Lane 2: His-tagged Mouse P-cadherin recombinant protein

Lysates/proteins at 20 ng/Lane.

Exposure time: 7 seconds; ECL: K1801;

4-20% 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 (HA723564) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21157_22.jpg Fig22: 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.
HAK21157 - Page 12
HAK21157_23.jpg Fig23: Fluorescence multiplex immunohistochemical analysis of mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Red), anti-Glucagon (ET1702-20, Green), anti-Insulin (ET1601-12, White), anti-CK19 (ET1601-6, Magenta) and anti-aSMA (ET1607-53, Yellow) on mouse pancreas. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1702-20 (1/6,000 dilution), ET1601-12 (1/8,000 dilution), ET1601-6 (1/5,000 dilution) and ET1607-53 (1/10,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HAK21157_24.jpg Fig24: Immunocytochemistry analysis of C6 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.
HAK21157 - Page 13
HAK21157_25.jpg Fig25: Application: IF-Tissue

Species: Mouse

Site: liver

Sample: Paraffin-embedded section

Antibody concentration: 1/200
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.