概述

  • 产品名称
    Anti-Hsp90抗体[D7a]
    参阅全部 Hsp90 一抗
  • 描述
    小鼠单克隆抗体[D7a] to Hsp90
  • 经测试应用
    适用于: WB, IP, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Chicken, Cow, Human, Pig
  • 免疫原

    Full length native protein (purified) corresponding to Chicken Hsp90 alpha aa 1-728. Full length protein HSP90 purified from chicken brain
    Database link: P11501

  • 阳性对照
    • HeLa (heat shocked) cell lysate; human Hsp90 alpha protein

性能

应用

Our Abpromise guarantee covers the use of ab59459 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
WB 1/500. Predicted molecular weight: 95 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

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

ICC/IF 1/200.

靶标

图片



  • Predicted band size : 95 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Hsp90 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hek 293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab59459 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab59459 was shown to specifically recognize Hsp90 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Hsp90 knockout cells were examined. Wild-type and Hsp90 knockout samples were subjected to SDS-PAGE. Ab59459 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

  • ab59459 staining Hsp90 alpha in the THP-1 monocyte/macrophage cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with Triton X-100 0.1% and blocked with 5% Serum for 60 minutes at 20°C. Samples were incubated with primary antibody (1/200 1% BSA/5% Goat Serum) for 1 hour at 20°C. An Alexa Fluor®647-conjugated Goat anti-mouse IgG polyclonal(1/500) was used as the secondary antibody.

    See Abreview

  • Anti-Hsp90 antibody [D7a] (ab59459) at 1/1000 dilution + Rat tissue lysates

    Predicted band size : 95 kDa
  • ab59459 staining Hsp90 alpha in human colon cancer tissue section by immunohistochemistry (Bouin's fixed paraffin embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave in citrate buffer. The primary antibody was used at dilution of 1/100,000 dilution using antibody amplifier™ system.

  • Overlay histogram showing HeLa cells stained with ab59459 (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 (ab59459, 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. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

文献

This product has been referenced in:
  • Perez-Branguli F  et al. Reverse Signaling by Semaphorin-6A Regulates Cellular Aggregation and Neuronal Morphology. PLoS One 11:e0158686 (2016). Read more (PubMed: 27392094) »
  • Morlé A  et al. Hyperthermia restores apoptosis induced by death receptors through aggregation-induced c-FLIP cytosolic depletion. Cell Death Dis 6:e1633 (2015). WB ; Human . Read more (PubMed: 25675293) »

See all 7 Publications for this product

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (4-15% SDS PAGE)
Loading amount
15 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

提交于 Apr 07 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (DU145)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
DU145
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

提交于 Mar 31 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (DU145)
Gel Running Conditions
Reduced Denaturing (4-15% SDS PAGE)
Loading amount
20 µg
Specification
DU145
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

提交于 Mar 31 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

提交于 Mar 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Uterus)
Specification
Uterus
Fixative
Formaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. Mukesh Jaiswal

Verified customer

提交于 Apr 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (THP-1 moncyte/macrophage cell line)
Specification
THP-1 moncyte/macrophage cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. James Harris

Verified customer

提交于 Aug 07 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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