Overview

  • Product name
    Anti-pan Cadherin antibody
    See all pan Cadherin primary antibodies
  • Description
    Rabbit polyclonal to pan Cadherin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, ICC, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chicken, Cow, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 850 to the C-terminus of Human pan Cadherin.

    Read Abcam's proprietary immunogen policy (Peptide available as ab17098.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; 3T3.

      This antibody gave a positive signal in the following tissue lysates: Mouse Heart Normal; Mouse Muscle normal; Human Heart Normal; Rat Heart Normal.

      This antibody gave a positive signal in the following cell lines: HeLa.

      This antibody gave a positive signal in the following tissues: Formalin Fixed Paraffin Embedded Human Liver Normal.

Properties

Applications

Our Abpromise guarantee covers the use of ab16505 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 2 µg/ml.

A diffuse signal is seen throughout the cells if higher concentrations are used (5-10µg/ml). We have had reports that the antibody works less well in this application in murine (3T3) cells. 

ICC Use a concentration of 2 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 135 kDa (predicted molecular weight: 100 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

Images

  • All lanes : Anti-pan Cadherin antibody (ab16505) at 1 µg/ml

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

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 100 kDa
    Observed band size: 125-140 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 40 kDa, 65 kDa, 75 kDa, 90 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute
  • ICC/IF image of ab16505 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16505, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • IHC image of pan Cadherin staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16505, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-pan Cadherin antibody (ab16505) at 1 µg/ml

    Lane 1 : Mouse heart
    Lane 2 : HeLa cell lysate
    Lane 3 : 3T3 cell lysate
    Lane 4 : Mouse muscle
    Lane 5 : Human heart
    Lane 6 : Mouse heart with Human pan Cadherin peptide (ab17098) at 1 µg/ml
    Lane 7 : HeLa cell lysate with Human pan Cadherin peptide (ab17098) at 1 µg/ml
    Lane 8 : 3T3 cell lysate with Human pan Cadherin peptide (ab17098) at 1 µg/ml
    Lane 9 : Mouse muscle with Human pan Cadherin peptide (ab17098) at 1 µg/ml
    Lane 10 : Human heart with Human pan Cadherin peptide (ab17098) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit conjugated to Alexafluor 680 at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 100 kDa

  • HeLa cells fixed in methanol and stained with ab16505 (2µg/ml). The cells were fixed in 100% methanol for 6 minutes at -20°C, then washed once in PBS. The 2 images show the cells stained with different secondary antibodies, Donkey anti Rabbit FITC (image A) and Donkey anti Rabbit Cy3 (image B). In each case ab16505 stains the plasma membrane.  In image A ab16505 is stained green and in image B ab16505 is stained red. In both images the DNA is stained with DAPI (blue).

  • Immunofluorescence analysis of HeLa cells, staining pan Cadherin (red) with ab16505.

    Cells were fixed with paraformaldehyde, permeabilized in methanol and blocked for 1 hour at room temperature in DPBS containing 2 mg/mL BSA, 1% fish gelatin, 0.1% Triton-X 100 and 5% goat serum. Cells were then incubated for 1 hour at room temperature with the primary antibody diluted in blocking buffer. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

References

This product has been referenced in:
  • Haas J  et al. Genomic structural variations lead to dysregulation of important coding and non-coding RNA species in dilated cardiomyopathy. EMBO Mol Med 10:107-120 (2018). WB ; Human . Read more (PubMed: 29138229) »
  • Webber HC  et al. The Role of Wnt/ß-Catenin Signaling and K-Cadherin in the Regulation of Intraocular Pressure. Invest Ophthalmol Vis Sci 59:1454-1466 (2018). ICC/IF ; Human . Read more (PubMed: 29625468) »
See all 43 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (liver tissue)
Permeabilization
No
Specification
liver tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Moon Han Chang

Verified customer

Submitted Jul 10 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human ovarian cystedenocarcinoma tissues)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: bicarbonate buffer pH 6.0
Permeabilization
No
Specification
human ovarian cystedenocarcinoma tissues
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 11 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample
Human Cell (ovarian carcinoma SKOV-3 cell line)
Specification
ovarian carcinoma SKOV-3 cell line
Permeabilization
Yes - 0.1% triton X for 5 min at RT
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Tissue sections (Day 9.5 embryo)
Specification
Day 9.5 embryo
Permeabilization
Yes - 0.1% saponin
Fixative
Paraformaldehyde

Dr. Nikhil Mittal

Verified customer

Submitted Sep 23 2013

Answer

Thank you for contacting us.

All of our pan-Cadherin antibodies have been tested in IHC-P with human samples and are guaranteed for this application and species.

We have 5 rabbit polyclonals: ab15186, ab45009, ab74635, ab6529, ab16505
and 2 mouse monoclonals: ab6528 and ab22744

Unfortunately, ab15186 comes as prediluted antibody. We do not have a concentrated version.

As mouse monoclonal, ab6528 would be a good choice - it has received 40 Abreviews (customer feedback with very good feedback for IHC-P) as well as 27 publications using the antibody.
As for a rabbit polyclonal that is not prediluted, I would recommend ab16505 based on Abreviews and publications.

We do not offer small sample sizes as this is not feasible for us operationally. Butwe guarantee all our products for TESTED species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your enquiry.

Below is a selection of antibodies that are suitable for detecting the plasma membrane and nuclear envelope. They all work in western blot and I would review the datasheets to confirm which species they are tested against.

Plasma membrane
ab4645 Anti-Pma1 antibody [40B7]
www.abcam.com/4645

ab7671 Anti-alpha 1 Sodium Potassium ATPase antibody [464.6]
www.abcam.com/ab7671

ab16505 Anti-pan Cadherin antibody
www.abcam.com/ab16505

ab22744 Anti-pan Cadherin antibody [mAbcam22744]
www.abcam.com/ab22744

Nuclear envelope
ab8980 Anti-Lamin A antibody [133A2]
www.abcam.com/ab8980

ab8984 Anti-Lamin A + C antibody [131C3]
www.abcam.com/ab8984

ab16048 Anti-Lamin B1 antibody
www.abcam.com/ab16048

I hope this is useful. Just let me know if you have further questions.

Read More

Answer

Thank you for bringing this topic to our attention.

I have spoken with the lab and they agree that the images do show some stronger staining clustered around the nucleus which does appear to be the golgi. Swiss prot actually states a golgi and cytoplamic vesicle localisation as the protein moves towards the cell membrane so this may explain it. However this product has been marked for continued testing.

Thank you again for contacting us. Please let me know if you have any questions.

Read More

Question
Answer

DISCOUNT CODE: *** Expiration date: February 28, 2012 I am very pleased to hear you would like to accept our offer and test ab16505 in IHC-Fr. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab16505 in IHC-Fr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.  

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293)
Specification
HEK293
Fixative
100% Ethanol
Permeabilization
No
Blocking step
(agent) for 30 minute(s) · Concentration: 0.1% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted May 21 2009

1-10 of 14 Abreviews or Q&A

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