Anti-MDM2 antibody [2A10]

• 产品名称

  Anti-MDM2抗体[2A10]
  参阅全部 MDM2 一抗

• 描述

  小鼠单克隆抗体[2A10] to MDM2

• 宿主

  Mouse

• 特异性

  Recognizes the ~90 kDa (apparent MW) MDM2 protein in A549 cells.

• 经测试应用

  适用于: ICC, WB, IHC-P, ICC/IF, Flow Cytmore details

• 种属反应性

  与反应: Human
  

• 免疫原

  Full length protein corresponding to MDM2 aa 250-350.
  Database link: Q00987-1

• 表位

  Within amino acids 294-339.

• 阳性对照

• A549 cell lysates. HeLa cells

• 常规说明

  This product was changed from ascites to tissue culture supernatant on 17 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at 4°C (stable for up to 12 months). Do Not Freeze.

• 存储溶液

  pH: 7.40
  Preservative: 0.05% Sodium azide
  Constituents: 0.88% Sodium chloride, Tris glycine

• 纯度

  Tissue culture supernatant

• 纯化说明

  Purified from TCS.

• 克隆

  单克隆

• 克隆编号

  2A10

• 同种型

  IgG2a

The Abpromise guarantee

Abpromise™承诺保证使用ab16895于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 4193 Human

• Omim: 164785 Human

• SwissProt: Q00987 Human

• Unigene: 484551 Human

• 别名

• Hdm 2 antibody

• Hdm2 antibody

• HDMX antibody

• MDM 2 antibody

• MDM2 antibody

• MDM2 oncogene E3 ubiquitin protein ligase antibody

• Mdm2 p53 E3 ubiquitin protein ligase homolog antibody

• Mdm2 transformed 3T3 cell double minute 2 p53 binding protein (mouse) binding protein 104kDa antibody

• MDM2_HUMAN antibody

• MDM2BP antibody

• Mouse Double Minute 2 antibody

• MTBP antibody

• Murine Double Minute Chromosome 2 antibody

• Oncoprotein Mdm2 antibody

• p53 Binding Protein Mdm2 antibody

• p53-binding protein Mdm2 antibody

• Ubiquitin protein ligase E3 Mdm2 antibody

• Ubiquitin protein ligase E3 Mdm2 antibody

• ACTFS antibody

• Double minute 2 protein antibody

• E3 ubiquitin-protein ligase Mdm2 antibody

• 

  Western blot - Anti-MDM2 antibody [2A10] (ab16895)

  Anti-MDM2 antibody [2A10] (ab16895) at 2 µg/ml + A549 whole cell lysate
  
  Predicted band size: 55 kDa
  
  
  
  

  Detection: chemiluminescence.

  This image was generated using the ascites version of the product.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 antibody [2A10] (ab16895)Image from Udeabor, Samuel Ebele et al. The Pan African Medical Journal 20 (2015): 140. doi:10.11604/pamj.2015.20. Fig 4. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of ameloblastoma tissue sections labeling MDM2 with ab16895. The sections were de-paraffinized, hydrated and then rinsed in phosphate-buffered solution (PBS). They were immersed in heat-induced epitope retrieval citrate buffer of concentration 15mMol and pH 6.0, diluted 1/10 with distilled water and incubated at 95ºC for 10 minutes. They were then placed in fresh citrate, cooled in water for 20 minutes and then rinsed in PBS for 6 minutes. Peroxidase blocking reagent was added to each section for 5 minutes, and the sections were rinsed in 0.1% PBS for 6 minutes. The specimen were incubated for 30 minutes with 1/100 dilution of Anti-MDM2 antibody [2A10] (ab16895), then rinsed with PBS, followed by incubation with undiluted HRP for 20 minutes. 1ml of diaminobenzidine solution was added to cover the specimen, followed by incubation in a humidity chamber for 15 minutes. The sections were then immersed in aqueous haematoxylin and rinsed in distilled water for 5 minutes. The tissue was dehydrated and subsequently rinsed with xylene. Distyrene plasticizer in xylene mounting fluid was then applied, and a cover slip placed. Hematoxylin and eosin staining.

  This image was generated using the ascites version of the product.

• 

  Flow Cytometry - Anti-MDM2 antibody [2A10] (ab16895)

  Overlay histogram showing HeLa cells stained with ab16895 (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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16895, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  

  This image was generated using the ascites version of the product.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [2A10] (ab16895)

  ab16895 staining MDM2 in MCF7 cells treated with progesterone (ab141252), by ICC/IF. Decrease in MDM2 expression correlates with increased concentration of progesterone, as described in literature.
  The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab141252 (progesterone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab16895 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  This image was generated using the ascites version of the product.

• 

  Western blot - Anti-MDM2 antibody [2A10] (ab16895)Image is courtesy of an anonymous Abreview

  Anti-MDM2 antibody [2A10] (ab16895) at 1/1000 dilution + Mouse Liver lysate at 40 µg with Milk, 2 hours at 25°C at 5 %
  
  Secondary
  Donkey anti-mouse IgG (HRP) at 1/10000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 55 kDa
  Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
  
  
  Exposure time: 1 minute
  
  
  

  This image was generated using the ascites version of the product.

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 antibody [2A10] (ab16895)Please note: for manual staining we recommend to optimize primary antibody concentration and incubation time (overnight incubation); amplification may be required.

  ab16895 staining Human normal tonsil. Staining is localised to nuclear + cytoplasmic compartments. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.

  Sections were stained using an automated system DAKO Autostainer Plus, at RT: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 min. They were then blocked with Dako Protein block for 10 min (containing casein 0.25% in PBS) , incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 min. Colorimetric detection was completed with DAB for 5 min. Slides were counterstained with Haematoxylin and coverslipped under DePeX.

  This image was generated using the ascites version of the product.

• 

  Immunocytochemistry - Anti-MDM2 antibody [2A10] (ab16895)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

  ab16895 (1/200) staining MDM2 in assynchronous HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). Please refer to Abreview for further experimental details.

  This image was generated using the ascites version of the product.

  See Abreview