- MA1-159 - Invitrogen Antibodies CDH2 antibody

Submitted references Adherens junctions stimulate and spatially guide integrin activation and extracellular matrix deposition.

Hadjisavva R, Anastasiou O, Ioannou PS, Zheltkova M, Skourides PA
Cell reports 2022 Jul 19;40(3):111091

Mechano-signalling, induced by fullerene C(60) nanofilms, arrests the cell cycle in the G2/M phase and decreases proliferation of liver cancer cells.

Sosnowska M, Kutwin M, Jaworski S, Strojny B, Wierzbicki M, Szczepaniak J, Łojkowski M, Święszkowski W, Bałaban J, Chwalibog A, Sawosz E
International journal of nanomedicine 2019;14:6197-6215

Diamond, graphite, and graphene oxide nanoparticles decrease migration and invasiveness in glioblastoma cell lines by impairing extracellular adhesion.

Wierzbicki M, Jaworski S, Kutwin M, Grodzik M, Strojny B, Kurantowicz N, Zdunek K, Chodun R, Chwalibog A, Sawosz E
International journal of nanomedicine 2017;12:7241-7254

Effect of N-cadherin misexpression by the mammary epithelium in mice.

Knudsen KA, Sauer C, Johnson KR, Wheelock MJ
Journal of cellular biochemistry 2005 Aug 15;95(6):1093-107

P-cadherin expression in breast carcinoma indicates poor survival.

Peralta Soler A, Knudsen KA, Salazar H, Han AC, Keshgegian AA
Cancer 1999 Oct 1;86(7):1263-72

P-cadherin expression in breast carcinoma indicates poor survival.

Peralta Soler A, Knudsen KA, Salazar H, Han AC, Keshgegian AA
Cancer 1999 Oct 1;86(7):1263-72

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on tissue extract (30 µg lysate) of Mouse Brain (Lane 1). The blot was probed with Anti-N-cadherin Monoclonal Antibody (Product # MA1-159, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 130 kDa band corresponding to N-cadherin was observed in the tissue tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of N-cadherin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR914282_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of N-cadherin was performed by loading 30 µg of SH-SY5Y Cas9 (Lane 1) andSH-SY5Y N-Cadherin KO (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-N-cadherin Monoclonal Antibody (13A9) (Product # MA1-159, 1:250 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to N-cadherin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of N-cadherin was performed by loading 25 µg of the indicated tissue lysate or whole cell lysates and 15 µL of PageRuler Prestained Protein Ladder (Product # 26616) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour. The membrane was probed with an N-cadherin monoclonal antibody (Product # MA1-159) at a concentration of 3 µg/mL overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of N-cadherin was performed using 90% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with N-cadherin Mouse Monoclonal Antibody (13A9) (Product # MA1-159) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of N-cadherin (green) in SH-SY5Y cells. Methanol-fixed cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with an N-cadherin monoclonal antibody (Product # MA1-159) at a dilution of 1:100 (right panel) or incubated in blocking buffer as a negative control (left panel) for 1 hour at room temperature, washed with PBS and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of N-cadherin was performed on HeLa, MCF7, and NIH/3T3 cells. Antigen-antibody complexes were formed by incubating 300 µg of HeLa, MCF7, and NIH/3T3 whole cell lysates with 2 µg of an N-cadherin monoclonal antibody (Product # MA1-159) overnight on a rocking platform at 4C. The immune complexes were captured on 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Eluted samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with StartingBlock T20 (Product # 37543) for 1 hour. The membrane was probed with an N-cadherin monoclonal antibody (Product # MA1-159) at a concentration of 2 µg/mL overnight rotating at 4C, washed in TBST, and probed with an HRP-conjugated anti-mouse light chain antibody at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 EGFR/AKT/mTOR and beta-catenin signaling in glioblastoma cells after treated with nanoparticles. Notes: ( A ) Confocal microscope images of U87 and U118 cells actin cytoskeleton. Cells were grown on extracellular matrix for 24 h and treated with diamond nanoparticles, graphite nanoparticles, or graphene oxide nanoparticles at a concentration of 20 mug/mL and incubated for 24 h. F-Actin was stained with phalloidin conjugated with Atto 633. ( B ) Western blot analysis of N-cadherin, pan-cadherin, vinculin, p-EGFR, and EGFR. GAPDH was used as a loading control. ( C ) Western blot analysis of nuclear and cytoplasmic protein fractions used for determination of beta-catenin protein level. PCNA and beta-tubulin were used as loading controls for nuclear and cytoplasmic fractions, respectively. ( D ) ELISA analysis of AKT and mTOR phosphorylation in comparison to control. Treatment with nanoparticles significantly reduced phospho-AKT ( P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Propidium iodide (PI) 488 assay analysis. Notes: Effect of C 60 on the number (percentage) of ( A ) HS-5, ( B ) HepG2 or ( C ) C3A cells in the cycle phases. ( D ) Relative HS-5, HepG2 and C3A cell population in G2/M cycle (%). ( E ) Western blot analysis of beta-catenin, N-cadherin, vinculin and PCNA. GAPDH was used as a loading control. Abbreviations: C 60 , fullerenes; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PCNA, proliferating cell nuclear antigen.

MA1-159

Antibody data