Anti-pan Cadherin抗体[CH-19] (ab6528)

概述

  • 产品名称Anti-pan Cadherin抗体[CH-19]
    参阅全部 pan Cadherin 一抗
  • 描述
    小鼠单克隆抗体[CH-19] to pan Cadherin
  • 经测试应用适用于: Flow Cyt, ICC/IF, IHC-P, WB, IHC-Fr, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Goat, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Human, Pig, Xenopus laevis, Sand rat, Snake, Zebrafish
  • 免疫原

    Synthetic peptide:

    DYDYLNDWGPRFKKLADMYGGGDD

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

  • 常规说明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.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt Use 1-2µg for 106 cells.
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.

靶标

Anti-pan Cadherin antibody [CH-19] 图像

  • 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 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

  • 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



  • Predicted band size : 125-140 kDa
    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 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

  • 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
    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

    This image was kindly supplied by Dr Vladimir Milenkovic by Abreview

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

    See Abreview

  • 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
    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?)

    This image is courtesy of an Abreview submitted by Dr. Vladimir Milenkovic

    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.

Anti-pan Cadherin antibody [CH-19] (ab6528)参考文献

This product has been referenced in:
  • Bender G  et al. Membrane-dependent Activities of Human 15-LOX-2 and Its Murine Counterpart: IMPLICATIONS FOR MURINE MODELS OF ATHEROSCLEROSIS. J Biol Chem 291:19413-24 (2016). Read more (PubMed: 27435673) »
  • Kulshrestha A  et al. Vacuolar ATPase 'a2' isoform exhibits distinct cell surface accumulation and modulates matrix metalloproteinase activity in ovarian cancer. Oncotarget 6:3797-810 (2015). IHC-P ; Human . Read more (PubMed: 25686833) »

See all 42 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (ovarian cancer cells)
Permeabilization Yes - 0.1% Triton-X (5min, RT).
Specification ovarian cancer cells
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Formaldehyde
Username

Dr. Gajendra Katara

Verified customer

提交于 Feb 15 2016

Application Western blot
Sample Human Cell lysate - whole cell (NERONAL CELL)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 50 µg
Treatment alpha synuclein
Specification NERONAL CELL
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Oct 05 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (Ovarian Carcinoma (TOV112D))
Specification Ovarian Carcinoma (TOV112D)
Permeabilization Yes - 0.1% Triton-x100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 18 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample Human Cell (HEK 293)
Specification HEK 293
Permeabilization Yes - 0.5% Triton X
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 07 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step Enzymatic - Buffer/Enzyme Used: 0.05% trypsin
Sample Mouse Tissue sections (lung)
Specification lung
Permeabilization Yes - 0.3% tritonX-100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 19 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Mouse Tissue sections (adult testes)
Specification adult testes
Permeabilization No
Fixative 4%PFA
Username

Dr. Qing Wen

Verified customer

提交于 May 01 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 37°C
Sample Rat Tissue sections (Vagus nerve)
Specification Vagus nerve
Permeabilization Yes - 0.2% triton X/PBS/Serum
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 28 2013

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 con...

Read More
Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS7 cell lysate - kidney)
Loading amount 4 µg
Specification COS7 cell lysate - kidney
Gel Running Conditions Reduced Denaturing (15% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Ms. Laura Fin

Verified customer

提交于 Aug 03 2012

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 unf...

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