Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker

• WB

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Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

All lanes:

Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952) at 1/500 dilution

Lane 1:

A431 cell lysate at 15 µL

Lane 2:

NIH-3T3 cell lysate at 15 µL

Lane 3:

HeLa cell lysate at 15 µL

Lane 4:

C6 cell lysate at 15 µL

Lane 5:

LNCap cell lysate at 15 µL

Secondary

All lanes:

HRP-Goat Anti-Mouse Igg (Fc) at 1/10000 dilution

Predicted band size: 99 kDa

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• Flow Cyt

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Flow Cytometry - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

Overlay histogram showing HeLa cells stained with ab98952 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab98952, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

AbReview41781****

Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

ab98952 staining N Cadherin in paraffin-embedded human lung cancer tissue by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with HOPE and blocked. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. A TRITC-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

Immunohistochemical analysis of paraffin-embedded Human lung cancer (1), colon cancer (2), ovarian cancer (3) and mammary cancer (4), using ab98952 at 1/200 dilution with DAB staining.

• IHC-P

AbReview42179****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

ab98952 staining N Cadherin in human NSCLC tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with using HOPE technique and blocked for 5 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

Immunofluorescence analysis of A431 cells using ab98952 at 1/200 dilution (green). Blue : DRAQ5 fluorescent DNA dye

• WB

CiteAb

Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

Western Blotting using Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker, ab98952. Publication image from Poutanen, M. et al., 2020, Nat Commun, 31959826. Legend direct from paper.

Androgen-deprivation upregulates SPINK1 in NE-transdifferentiated PCa cells.a Bar graph showing SPINK1 expression (top) and heatmap of AR-signaling associated genes including SPINK1 in long-term androgen deprived (AD) LNCaP cells (GSE8702). b Representative phase-contrast images of androgen-deprived LNCaP cells (LNCaP-AI). Red arrow-heads indicate neurite outgrowth. c QPCR data showing relative expression of SPINK1 and KLK3 using same cells as b. d Immunoblot assay for SPINK1, PSA, SYP, ENO2, SOX2 and REST using same cells as in b. e Immunostaining for SPINK1 using same cells as in b. f Schematic representation of NE-transdifferentiation (NE-td) using doxycycline (dox)-inducible AR-V7 overexpressing LNCaP cells subjected to androgen deprivation (AD) with or without induction (40 ng/ml) at day 10 and cultured upto 30 days. g QPCR data showing relative expression of SPINK1 and KLK3 using same cells as f. h Immunoblot assay for AR, AR-V7, SPINK1, and PSA using same cells as f. i QPCR data showing relative expression of SYP, NCAM1, ENO2 and CHGA using the same cells as f. j Schema describing generation of LNCaP-AI-shSPINK1 and LNCaP-AI-shSCRM cells by subjecting stable LNCaP-shSPINK1 and LNCaP-shSCRM cells to androgen deprivation (AD) for 30 days. k Representative images for the neurite outgrowths in LNCaP-AI-shSCRM cells and LNCaP-AI-shSPINK1 as j (top). QPCR data showing relative expression of SPINK1 (bottom, left) and measurement of neurite outgrowth (bottom, right). l Immunoblot analysis for SPINK1, E-Cad, VIM, and N-Cad expression using same cells as j. m Same as in l, except phospho (p) and total (t) AKT, SYP and ENO2 expression. n Representative IHC images for SPINK1 in SPINK1-negative (SPINK1−, top) and SPINK1-positive (SPINK1+, bottom) PCa tumor cores of the VPC tissue microarray (scale bar = 200 µm). Bar plot showing percentage cases of SPINK1 in untreated (n = 33) and neoadjuvant-hormone therapy (NHT) treated patients (n = 55). Experiments were performed with n = 3 biologically independent samples; data represents mean ± SEM. For panels b, e, k scale bar represents 20 µm. For panel c two-way ANOVA, Dunnett’s multiple-comparisons; g, i two-way ANOVA, Sidak’s multiple-comparisons test; k one-way ANOVA, Dunnett’s multiple-comparisons test were applied. ∗P ≤ 0.05 and ∗∗P ≤ 0.001. Source data for d, h, l, m are provided as a Source Data file.

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• WB

CiteAb

Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (AB98952)

Western Blotting using Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker, ab98952. Publication image from Luo, J. et al., 2020, Theranostics, 32089740. Legend direct from paper.

ZNF471 inhibits ESCC cell migration and invasion and EMT. (A) Morphological changes of ESCC cells infected with control vector and ZNF471 by phase contrast microscopy. Scale bars : 200 µm. (B) Cell migration abilities of ESCC cells evaluated by wound healing assays. Photographs captured at 0, 12 and 24 h. The cellular migration (C) and invasion (D) abilities of ESCC cells upon ectopic expression of ZNF471 were measured by Transwell assays with or without Matrigel. Representative images were photographed following fixation and staining. Scale bars : 50 µm. (E, F) qRT-PCR and western blot analysis of EMT, and downstream target markers. Student's test was used. Data are presented as the mean ± SD. *p<0.05, **p<0.01, ***p<0.001.

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