Anti-pan Cadherin 抗体 [mAbcam22744] - Plasma Membrane Marker (ab22744)

製品の概要

  • 製品名Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker
    pan Cadherin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [mAbcam22744] to pan Cadherin - Plasma Membrane Marker
  • 特異性Detects a weaker band in human heart than in rat heart.
  • アプリケーション適用あり: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human, African Green Monkey
    交差が予測される動物種: Chicken, Dog, Xenopus laevis, Zebrafish
  • 免疫原

    Synthetic peptide at the C-terminus of Human Cadherins.

    .

  • 特記事項

    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)

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker 画像

  • Lane 1 : Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker (ab22744) at 1 µg/ml (Blocked in 5% BSA)
    Lane 2 : Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker (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
  • All lanes : Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker (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
  • 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).

  • 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).

  • Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker (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

  • 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

  • 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.

Anti-pan Cadherin antibody [mAbcam22744] - Plasma Membrane Marker (ab22744) 使用論文

This product has been referenced in:
  • Li J  et al. Anti-KCNQ1 K+ channel autoantibodies increase IKs current and are associated with QT interval shortening in dilated cardiomyopathy. Cardiovasc Res 98:496-503 (2013). Read more (PubMed: 23447643) »
  • Izumi H & Kaneko Y Evidence of asymmetric cell division and centrosome inheritance in human neuroblastoma cells. Proc Natl Acad Sci U S A 109:18048-53 (2012). ICC/IF ; Human . Read more (PubMed: 23064640) »

See all 6 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

投稿 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

投稿 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

投稿 Jul 15 2015

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

Abcam user community

Verified customer

投稿 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

投稿 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

投稿 Dec 08 2011

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