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Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

Reviews (13)Q&A (6)References (18)
Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
  • Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
  • Western blot - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

Key features and details

  • Mouse monoclonal [mAbcam22744] to pan Cadherin
  • Suitable for: ICC/IF, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

HRP

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: ICC/IF, WBmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Dog, Xenopus laevis, Monkey, Zebrafish, African green monkey

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • General notes

    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.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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
ICC/IF (4)
Use a concentration of 5 µg/ml.
WB (6)
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.
Notes
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.
Application notes
Is unsuitable for Flow Cyt.

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).

  • 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 (18)

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

ab22744 has been referenced in 18 publications.

  • Kong LR  et al. A common MET polymorphism harnesses HER2 signaling to drive aggressive squamous cell carcinoma. Nat Commun 11:1556 (2020). PubMed: 32214092
  • Joksimovic SL  et al. Selective inhibition of CaV3.2 channels reverses hyperexcitability of peripheral nociceptors and alleviates postsurgical pain. Sci Signal 11:N/A (2018). PubMed: 30154101
  • Storman EM  et al. Physical Linkage of Estrogen Receptor a and Aromatase in Rat: Oligocrine and Endocrine Actions of CNS-Produced Estrogens. Endocrinology 159:2683-2697 (2018). PubMed: 29771302
  • 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). 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). PubMed: 28751455
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 19 Abreviews or Q&A

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

 Excellent
Abreviews
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

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

 Poor
Abreviews
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

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/pan-Cadherin-antibody-mAbcam22744-ab22744.html&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

1-10 of 19 Abreviews or Q&A

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