Overview

  • Product name
    Anti-pan Cadherin antibody [CH-19]
    See all pan Cadherin primary antibodies
  • Description
    Mouse monoclonal [CH-19] to pan Cadherin
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, WB, IHC-Fr, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Goat, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Human, Pig, Xenopus laevis, Sand rat, Snake, Zebrafish
  • Immunogen

    Synthetic peptide:

    DYDYLNDWGPRFKKLADMYGGGDD

    conjugated to KLH, corresponding to amino acids 889-912 of Chicken N-Cadherin (cdh2).

  • General notes

    This antibody can be used as a marker for the plasma membrane in cells which express cadherins - see reviews.

    This product is useful for the detection of members of the cadherin family or genetically engineered proteins containing the C-terminal cadherin tail, and for demonstration of adherens type cell-cell junctions regardless of their cadherin type.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab6528 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
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 125-140 kDa.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

Target

Images

  • Cellular localization: cell membrane. Specificity This antibody reacts with pan cadherin with a distinct 135 kDa band from a wide variety of tissues. Cadherins are members of a multigene family of single chain glycoprotein receptors mediating Ca2+ dependent cell-cell adhesion

  • All lanes : Anti-pan Cadherin antibody [CH-19] (ab6528) at 1/2000 dilution

    Lane 1 : Porcine kidney extract - whole cell lysate
    Lane 2 : Porcine RPE cells - whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : An HRP-conjugated goat polyclonal to mouse IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 125-140 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?

    See Abreview

  • Immunofluorescent imaging of human cells (U2OS) with ab6528
    reveals the expected highly specific membranous distribution.

    IF was performed with a standard paraformaldehyde technique (fixed in
    PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS
    for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.
    Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour,
    secondary antibody for 30 minutes.  All blocking and incubation steps
    carried out at 37 degrees C.

  • ab6528 (1/1500) positively staining cadehrin in mouse kidney tissue sections, as secondary antibody goat anti mouse biotin conjugated was used. Please see accompanying abreview for additional information.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab6528 (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 (ab6528, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.

  • Madin Darby Bovine Kidney (MDBK) extract was separated on SDS-PAGE and probed with ab6528 at 1:2000. The antibody was developed with HRP Mouse Fab ads/HIgG conjugate and a NBT/BCIP substrate.
  • Ab6528 positively staining formaldehyde fixed HEK 293 cells at 1/200 in conjunction with goat anti mouse (Alexa 546) 1/1500.

    This image is an edited version of an image received courtesy of an Abreview submitted by Kun Liu on 19 September 2005. We do not have any further information relating to this image.

    See Abreview

  • All lanes : Anti-pan Cadherin antibody [CH-19] (ab6528) at 1/2000 dilution

    Lane 1 : ARPE-19 cells, membrane fraction
    Lane 2 : HEK 293 cells, membrane fraction

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat anti-mouse conjugated to HRP at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 125-140 kDa
    Observed band size: 135 kDa why is the actual band size different from the predicted?
    Additional bands at: 60 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 10 minutes


    Gel running conditions are denaturing and 10% SDS.
    Blocked using 5% milk for 30 minutes at 25°C.

    See Abreview

  • ab6528 staining pan Cadherin in mouse Cor1 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with formaldehyde, permeabilized with TBS, BSA, azide and Triton and blocking with 1% BSA  for 30 minutes at RT was performed. Samples were incubated with primary antibody (1/1500: in TBS, BSA, azide and 0.3%Triton) for 2 hours. An Alexa Fluor®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/1000 dilution. The image was captured using Z-stacks/Zeiss Apotome microscope rig.

    See Abreview

References

This product has been referenced in:
  • Schrade K  et al. An AKAP-Lbc-RhoA interaction inhibitor promotes the translocation of aquaporin-2 to the plasma membrane of renal collecting duct principal cells. PLoS One 13:e0191423 (2018). Read more (PubMed: 29373579) »
  • Boutin ME  et al. A three-dimensional neural spheroid model for capillary-like network formation. J Neurosci Methods 299:55-63 (2018). IHC-P . Read more (PubMed: 28143748) »
See all 67 Publications for this product

Customer reviews and Q&As

1-10 of 22 Q&A

Answer

The pan-cadherin antibody ab6528 is raised against a cytoplasmic sequence of chicken cadherin 2 (CDH2) which is conserved in Chinese hamster cadherins. So, although it will be capable of detecting CHO cadherins, detection in cells will require fixation and permeabilization, as the antibody needs to cross the plasma membrane to reach the intracellular sequence.

Read More

Answer

This antibody reacts extensively with all known members of the cadherin family. The cadherins migrate in the molecular weight range of 125 to 140 kDa and are tissue dependent. Therefore, Pan-Cadherin detects cadherin which have the carboxy terminal conserved among the cadherin family proteins. Some examples of such Cadherin include E-Cadherin, N-Cadherin and P-Cadherin.

Read More

Answer

Thank you for contacting us and your interest in our products.

I would suggest that using a plasmamembrane dye or antibody capable of detecting markers of the membrane would work well in what you are trying to achieve.

We do not unfortunately have any cell membrane dyes currently.

However, you could use an antibody directed against a plasma membrane target, such as pan cadherin. I would suggest having a look at the antibody https://www.abcam.com/pan-Cadherin-antibody-CH-19-ab6528.html.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

Read More

Answer

That sounds very good.

Please do let me know if you have more questions or are satisfied or not satisfied with any of our products.

Read More

Answer

Thank you for your reply.
I assume you are refering to ab6528. This antibody has been used in 27 publications. The reviews are submitted by customers and the results may vary depending on samples used, species used as well as protocols used. Also, the WB images show mainly one strong dominating band, while some other bands seen can be accounted for heavy chains of endogenous IgG in case of tissue samples. WithMDBK cellsa single band was obtained.

In any case, we guarantee the antibody for theapplications and species listed as tested 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.

Thus, we stand behind our product, but should you experience any problems, we are happy to troubleshoot and replace, credit or refund the antibody should it not perform to your satisfaction.

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 reply.

Yes, although the datasheet states that enzymatic antigen retrieval should be used, at least 2customers publishedAbreviews were citrate buffer was employed and it worked well, too.
for human: https://www.abcam.com/index.html?datasheet=6528&tab=abreviews&intabreviewid=10495
for mouse: https://www.abcam.com/index.html?datasheet=6528&tab=abreviews&intabreviewid=7553

Thus, citrate buffer would be a suitable alternative antigen retrieval method.

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

Vielen Dank für Ihren Anruf. Es tut mir leid, dass Sie Probleme mit diesem beuen Röhrchen von Antikörper hatten. Wie am Telefon besprochen, habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer xx und sollte in der ersten Januarwoche bei Ihnen ankommen. Ich freue mich wieder von Ihnen zu hören, wie die neue Charge funktioniert. Ich wünsche Ihnen eine wunderbare Weihnachtszeit und einen guten Rutsch in das neue Jahr!

Read More

Question
Answer

Thanks for your call today and for letting us know about the trouble with ab6528. As we discussed, I am sending a vial of ab6528 free of charge on the order ***. This vial is in stock and should arrive tomorrow morning. Please keep me updated about how this replacement antibody works for you, and let me know if there is anything else that we can do for you.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 998881. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting us. I recommend ab7671, Anti-alpha 1 Sodium Potassium ATPase antibody: Click here (or use the following: https://www.abcam.com/index.html?datasheet=7671). The datasheet includes two images of sub-confluent HEK293 immunocytochemistry, as well as a western blot of HEK293 lysate. For cadherin, I suggest ab6528. The datasheet for this antibody includes an image of HEK293 ICC (red stain, DAPI counterstain) submitted by a customer in an Abreview, but the cells are in a clump of two or three, and not completely isolated from one another: Click here (or use the following: https://www.abcam.com/index.html?datasheet=6528). It is difficult to choose between the two. Both have been published many times and both are well-reviewed by our customers. Based on western blot data, the ATPase antibody appears to be somewhat more specific for its target than the cadherin antibody. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

1-10 of 22 Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up