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.

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

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

  • 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

  • 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 (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

  • 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.



  • 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 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

  • 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

  • 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

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

This product has been referenced in:
  • Haghani A  et al. Edible bird's nest modulate intracellular molecular pathways of influenza A virus infected cells. BMC Complement Altern Med 17:22 (2017). Read more (PubMed: 28056926) »
  • Serrano-Pozo A  et al. Acute and Chronic Sustained Hypoxia Do Not Substantially Regulate Amyloid-ß Peptide Generation In Vivo. PLoS One 12:e0170345 (2017). WB ; Mouse . Read more (PubMed: 28099462) »

See all 46 Publications for this product

Product Wall

The pan-cadherin antibody ab6528 is raised against a cytoplasmic sequence of chicken cadherin 2 (CDH2) which is conserved in Chinese hamster cadherins. So, although it will be capable of detecting CHO cadherins, detection in cells will require fixation...

Read More
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

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