Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

Overview

  • Product name
    Anti-pan Cadherin antibody [mAbcam22744]
    See all pan Cadherin primary antibodies
  • Description
    Mouse monoclonal [mAbcam22744] to pan Cadherin
  • 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.

    .

  • 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.

     

    Alternative versions available:

    Anti-pan Cadherin antibody (HRP) [mAbcam22744] - Plasma Membrane Marker (ab197725)

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

Anti-pan Cadherin antibody [mAbcam22744] images

  • 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)

    Lane 1 : Heart (Rat) Tissue Lysate
    Lane 2 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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
  • 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).

  • 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.

  • 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
    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
  • 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

  • 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

    This image is courtesy of an Anonymous abreview.

    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

  • 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 for Anti-pan Cadherin antibody [mAbcam22744] (ab22744)

This product has been referenced in:
  • Xia Z  et al. Zebrafish slc30a10 deficiency revealed a novel compensatory mechanism of Atp2c1 in maintaining manganese homeostasis. PLoS Genet 13:e1006892 (2017). ICC/IF ; Zebrafish . Read more (PubMed: 28692648) »
  • Deng Q  et al. Androgen Receptor Localizes to Plasma Membrane by Binding to Caveolin-1 in Mouse Sertoli Cells. Int J Endocrinol 2017:3985916 (2017). WB, IP, IF ; Mouse . Read more (PubMed: 28642789) »

See all 12 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Junmo Hwang

Verified customer

Submitted Jun 28 2016

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
Username

Dr. Junmo Hwang

Verified customer

Submitted Mar 11 2016

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
Username

Abcam user community

Verified customer

Submitted Jul 15 2015

Application
IHC - Wholemount
Sample
Zebrafish Embryo (whole embryo staining)
Specification
whole embryo staining
Username

Abcam user community

Verified customer

Submitted Jun 10 2014

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
Username

Abcam user community

Verified customer

Submitted Dec 06 2012

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 ...

Read More

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...

Read More

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

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:

http://www...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Dec 08 2011

1-10 of 19 Abreviews or Q&A

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