Anti-GRP94抗体[9G10] (ab90458)

概述

  • 产品名称Anti-GRP94抗体[9G10]
    参阅全部 GRP94 一抗
  • 描述
    大鼠单克隆抗体[9G10] to GRP94
  • 经测试应用适用于: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Xenopus laevis, Monkey
  • 免疫原

    Native chicken GRP94 protein

  • 阳性对照
    • Recombinant GRP94 protein HeLa, Heat shocked HeLa, Mouse liver or Vero cell lysate IHC-P: human pancreas FFPE tissue sections IF/ICC: HeLa cell line.

性能

应用

Our Abpromise guarantee covers the use of ab90458 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 1/1000. Predicted molecular weight: 92 kDa.
IP 1/100.
Flow Cyt 1/100. ab18450-Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

靶标

Anti-GRP94 antibody [9G10] 图像

  • ICC/IF image of ab90458 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab90458, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96887, DyLight® 488 goat anti-rat IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM

  • All lanes : Anti-GRP94 antibody [9G10] (ab90458) at 1/1000 dilution

    Lane 1 : GRP94 recombinant protein at 0.1 µg
    Lane 2 : HeLa cell lysate at 20 µg
    Lane 3 : Mouse liver lysate at 20 µg
    Lane 4 : Vero cell lysate at 20 µg


    Predicted band size : 92 kDa
  • Overlay histogram showing HeLa cells stained with ab90458 (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 (ab90458, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • IHC image of GRP94 staining in human pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab90458, 5µg/ml, for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-GRP94 antibody [9G10] (ab90458)参考文献

ab90458 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample Sacrophilus harrisii (Tasmanian Devil) Cell lysate - whole cell (C5065 (tasmanian devil cell line) K562 (human))
Gel Running Conditions Reduced Denaturing
Loading amount 30 µg
Specification C5065 (tasmanian devil cell line) K562 (human)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 Jun 05 2015

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