Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

Reviews (13)Q&A (6)References (15)

Overview

  • Product name
    Anti-pan Cadherin antibody [mAbcam22744]
    See all pan Cadherin primary antibodies
  • Description
    Mouse monoclonal [mAbcam22744] to pan Cadherin
  • Host species
    Mouse
  • Specificity
    Detects a weaker band in human heart than in rat heart.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Chicken, Dog, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide at the C-terminus of Human Cadherins.

    Read Abcam's proprietary immunogen policy .

  • General notes

    This antibody clone is manufactured by Abcam.

    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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab22744 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 1µg for 106 cells.

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

 
ICC/IF Use a concentration of 5 µg/ml.
WB 1/1000. Detects a band of approximately 125-140 kDa (predicted molecular weight: 125 kDa).

Abcam recommends using 3-5% milk as the blocking agent. Please see Western Blot data below.
IHC-P 1/2000.

Target

Images

  • Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Lane 1 : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml (Blocked in 5% BSA)
    Lane 2 : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml (Blocked in 5% Milk)

    All lanes : Heart (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 125 kDa


    Exposure time: 30 seconds
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

    ICC/IF image of ab22744 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22744, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 phalloidin was used to label F-actin (red).

  • Flow Cytometry - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Flow Cytometry - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Overlay histogram showing HEK293 cells stained with ab22744 (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 (ab22744, 1µg/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.

  • Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    All lanes : Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1 µg/ml

    Lane 1 : Heart (Rat) Tissue Lysate (blocked with 5% Milk)
    Lane 2 : Heart (Mouse) Tissue Lysate (blocked with 5% Milk)
    Lane 3 : Heart (Human) Tissue Lysate - adult normal tissue (blocked with 5% Milk)
    Lane 4 : Heart (Rat) Tissue Lysate (blocked with 3% Milk)
    Lane 5 : Heart (Mouse) Tissue Lysate (blocked with 3% Milk)
    Lane 6 : Heart (Human) Tissue Lysate - adult normal tissue (blocked with 3% Milk)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 125 kDa
    Observed band size: 125 kDa
    Additional bands at: 25 kDa, 58 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)Image courtesy of an anonymous Abreview.
    ab22744 staining pan Cadherin in mixed glia prepared from mouse brain by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol, permeabilised in 0.5% (w/v) saponin and then blocked using 10% serum for 2 hours at 23°C. Samples were then incubated with primary antibody at 1/100 for 2 hours at 23°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 488 (green) used at a 1/400 dilution. Counterstained with DAPI (blue).

    See Abreview

  • Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)This image is courtesy of an Anonymous abreview.
    Anti-pan Cadherin antibody [mAbcam22744] (ab22744) at 1/500 dilution + Mouse cultured cortical neurons at 20 µg

    Secondary
    HRP-conjugated Goat Anti-Mouse IgG (H+L) polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 125 kDa


    Exposure time: 1 minute


    Blocking performed with 5% milk for 1 hour.
    Primary diluted with PBS + 0.5% Tween20 and incubated for 12 hours at 4°C
    Performed under denaturing conditions.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
    Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

    ICC/IF image of ab22744 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22744, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 phalloidin was used to label F-actin (red).

References

This product has been referenced in:
  • Deng Q  et al. Vesicle-Associated Membrane Protein-Associated Protein A Is Involved in Androgen Receptor Trafficking in Mouse Sertoli Cells. Int J Endocrinol 2018:4537214 (2018). Read more (PubMed: 29686703) »
  • Zemel BM  et al. Calcineurin Dysregulation Underlies Spinal Cord Injury-Induced K+ Channel Dysfunction in DRG Neurons. J Neurosci 37:8256-8272 (2017). Read more (PubMed: 28751455) »
See all 15 Publications for this product

Publishing research using ab22744? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 19 Abreviews or Q&A

Western blot abreview for Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (7%)
Loading amount
10 µg
Specification
HEK293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Read More

Dr. Junmo Hwang

Verified customer

Submitted Jun 28 2016

Western blot abreview for Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (Cortex)
Gel Running Conditions
Reduced Denaturing (7%)
Loading amount
30 µg
Specification
Cortex
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Read More

Dr. Junmo Hwang

Verified customer

Submitted Mar 11 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (J774A.1 macrophages)
Permeabilization
Yes - 0.05% saponin
Specification
J774A.1 macrophages
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Methanol
Read More

Abcam user community

Verified customer

Submitted Jul 15 2015

IHC - Wholemount abreview for Anti-pan Cadherin [mAbcam22744] antibody - Plasma Membrane Marker

 Poor
Abreviews
Application
IHC - Wholemount
Sample
Zebrafish Embryo (whole embryo staining)
Specification
whole embryo staining
Read More

Abcam user community

Verified customer

Submitted Jun 10 2014

Western blot abreview for Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker

 Average
Abreviews
abreview image
Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Dec 06 2012

Question

Product code: 22744
Lot number: GR64651-1

Inquiry: Problem: Multiple bands detected (see attached file) Sample: Human colon adenocarcinoma cell line HCT-116 Specification: Whole cell lysate Amount of protein loaded: 10 ug Electrophoresis conditions: Reducing and denaturing 10% SDS-PAGE Blocking conditions: 5% milk in TBS-T for 2 hr, room temperature Primary antibody conditions: 1:5000 in 2% BSA in TBS-T, 2 hr, room temperature Secondary antibody: Non-Abcam Sheep anti-mouse HRP Secondary antibody conditions: 1:5000 in 2% BSA in TBS-T, 1 hr, room temperature Detection: DURA (Pierce), 3 min Wash steps: 3 x 5 min using TBS-T

Read More
Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab22744 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab22744.

1.) I suggest to not use Tween in the blocking solution. The detergent could mask proteins that should be blocked and therefore allow the background bands. Other users have good experience with blocking over night .

2.) To achieve the most specific binding of the antibody I recommend to incubate it at 4C over night.

3.) Maybe the addition of DTT was reducing agent instead of beta mercaptoethanol will help reduce the background..

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Question

Hi, Could you recommend a good antibody that I could use as loading control for western blot on cell membrane fractions? Thanks

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

Question

Thanks for your prompt reply. I will use methanol to fix my cells and try cadherin antibody again. I will let you know the results in a few days.

Read More
Answer

OK, good luck. If it does not work, I will have some suggestions for a different membrane marker, most likely anti-Na/K ATPase.

Read More

Question

I have tested the new cadherin antibody (ab22744). It works better than the previous one. However, it is not what I expected to be. Here attached the images.

I followed the suggested concentration (5 ug/ml) and general protocol for immunocytochemistry. I also used a higher concentration (10 ug/ml). The problem is not because of the cadherin concentration but the specificity to the cadherin as a plasma membrane marker within my Sol8 cells (a myogenic cell line).

I am thinking of using another plasma membrane marker. How about the Na/K ATPase? I wish I can get another replacement.

I will look forward to your reply. Thanks for your time and help.

Read More
Answer

Thank you for the images of the staining with ab22744.

I agree the staining is not what is usually observed for cadherin. I think you are expecting something like what one of our reviewers saw. The data is at this link:

https://www.abcam.com/index.html?datasheet=22744&tab=abreviews&intabreviewid=22768

Are you still fixing your cells with PFA? It may be that correct staining is dependent on the fixation protocol. The data we have for ab22744 is all from methanol-fixed cells. I suggest trying this if you are able to, fixation in cold methanol for five minutes, followed by rinsing with PBS.

Please let me know if you are willing to try this. If not, we can discuss replacement with a Na ATPase antibody.

Read More

Immunohistochemistry (Frozen sections) abreview for Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker

 Poor
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Apteronotus leptorhynchus Tissue sections (Brain, Spinal cord)
Specification
Brain, Spinal cord
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X-100
Blocking step
3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Read More

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Dec 08 2011

1-10 of 19 Abreviews or Q&A

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