The largest database of trusted experimental protocols

4 protocols using rabbit anti n cadherin

1

Immunohistochemical Analysis of Tissue Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were collected at the designated times and snap-frozen in optimal cutting temperature (OCT) medium (Sakura Finetek, Japan), and 8-μm sections were prepared and stained with the following antibodies; Rabbit anti-fibronectin (1:100) (Millipore, Temecula, CA, USA), rabbit anti-vimentin (1:100) (Cell Signalling Technology, Danvers, MA, USA), rabbit anti-N-cadherin (1:1000) (Thermo Scientific, Rockford, IL, USA), as described previously13 (link),41 (link). Stained sections were analysed using fluorescent or bright-field imaging microscopy (Leica, Germany) and ImagePro Plus Capture and Analysis software (Media Cybernetics, Rockville, MD). Vimentin, Fibronectin and N-Cadherin positive areas were quantified in 15 independent fields/section using Image Pro-Plus software41 (link),44 (link).
+ Open protocol
+ Expand
2

Inhibitor and Antibody Validation for Cell Studies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Inhibitors were as follows: CinnGEL-2ME and Gö-6976 from Biomol, apocynin from Acros Organics. Inhibitors and antibody crosslinking of VCAM-1 did not affect cell viability during the time courses of these studies, consistent with our previous reports [1 (link), 24 (link), 25 (link)]. Antibodies were as follows: Rat anti-mouse VCAM-1 (clone MVCAM.A), mouse anti-human VCAM-1 (clone 51-10C9), rat anti-mouse VE-Cadherin (CD144, cat#550548), rat IgG (isotype antibody, clone R35-95), and FITC-conjugated goat anti-rabbit Ig (cat#554020) [PharMingen]; zenon Alexa Fluor 568-labeled (cat#Z25106, Molecular Probes) rat anti-mouse CD45 (clone I3/2.3), goat anti-mouse IgG1 (cat#1070-01) and goat anti-rat IgG (cat#3050-01) [Southern Biotech]; mouse anti-phosphotyrosine (cat#9411) [Cell Signaling Technology]; Texas Red-conjugated goat anti-rat Ig (cat#R40005) [Caltag]; rabbit anti-human ZO-1 (cat# 41090971), FITC-conjugated rabbit anti-ZO-1 (cat#bs-1329R-FITC)[Bioss], and rabbit anti-phosphoserine (cat#61-8100) [Zymed Lab, Inc]; Alexa 594-conjugated donkey anti rabbit N-Cadherin (cat#150080) [Abcam]; rabbit anti-N-Cadherin (cat#PA5-19486)[Thermo Scientific]; rabbit anti-N-terminal region of angiomotin (cat# ARP34660_P050)[Aviva Systems Biology]; HRP-conjugated donkey anti-rabbit antibody [Amersham Pharmacia]; HRP-conjugated goat anti-mouse IgG [Bio-Rad].
+ Open protocol
+ Expand
3

Immunohistochemical Analysis of Tumor Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tumour and lung tissues were harvested and snap-frozen in optimal cutting temperature (OCT) medium (Sakura Finetek, Torrance, CA, USA), and 8-μm sections were prepared. Tumour sections were stained with the following antibodies: rabbit anti-fibronectin (Millipore, Temecula, CA, USA), rabbit anti-vimentin (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-N-cadherin (Thermo Scientific, Rockford, IL, USA), followed by Alexa 488 or Alexa 568-conjugated goat anti-rabbit IgG. Lung sections were stained with hematoxylin (Vector Laboratories, Burlingame, CA, USA) and eosin (Fisher Scientific Co., Kalamazoo, MI, USA) following the manufacturer’s protocol. We used fluorescent or bright-field imaging microscopy (Leica, Wetzlar, Germany) and ImagePro Plus Capture and Analysis software (Media Cybernetics, Rockville, MD, USA) to analyse the stained sections. We used Image Pro-Plus software to quantify stained areas from 10–15 independent fields/sections.
+ Open protocol
+ Expand
4

Immunofluorescence Staining of Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were cultured in 8-well chamber glass slides coated with 0.1% gelatin until confluence. They were fixed with 4% paraformaldehyde, rinsed with PBS, and blocked with 3% serum and 1% BSA for immunostaining using following primary antibodies: rabbit anti-ZEB1 (gift from Dr. Douglas Darling, 1:1000)35 (link), mouse anti-Ki67 (BD Pharmingen, Cat# 550609, 1:50), mouse anti-E-cadherin (BD Biosciences, Cat# 610181), rabbit anti-N-cadherin (ThermoFisher, Cat# PA5-19486) together with an anti-rabbit IgG secondary antibody conjugated with Alexa fluor 568 (Invitrogen, Cat# A11011, 1:500), or an anti-mouse IgG second antibody conjugated with Alexa fluor 488 (Invitrogen, Cat# A11001, 1:500). Nuclei were counterstained with Hoechst dye (Invitrogen, Cat# H1399, 1:500), and images were captured by an invert fluorescent microscope.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!