Anti-SMN/Gemin 1 antibody [2B1]

• 产品名称

  Anti-SMN/Gemin 1抗体[2B1]
  参阅全部 SMN/Gemin 1 一抗

• 描述

  小鼠单克隆抗体[2B1] to SMN/Gemin 1

• 宿主

  Mouse

• 经测试应用

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

• 种属反应性

  与反应: Mouse, Human, Xenopus laevis
  

• 免疫原

  Recombinant full length protein corresponding to Human SMN/Gemin 1.
  Database link: Q16637

• 常规说明

  
  

  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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  Preservative: 0.1% Sodium azide
  Constituent: PBS

• 纯度

  Protein A purified

• 纯化说明

  Purified from tissue culture supernatant.

• 克隆

  单克隆

• 克隆编号

  2B1

• 骨髓瘤

  Sp2/0

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab5831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: 6606 Human

• Entrez Gene: 6607 Human

• Entrez Gene: 20595 Mouse

• Omim: 600354 Human

• SwissProt: Q16637 Human

• SwissProt: P97801 Mouse

• Unigene: 202179 Human

• Unigene: 535788 Human

• Unigene: 2025 Mouse

• 别名

  hide

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• Gemin 1 antibody

• 

  Immunocytochemistry/ Immunofluorescence - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)

  ICC/IF image of ab5831 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab5831, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.
  
  

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)

  IHC image of ab5831 staining in human normal cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab5831, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer 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.
  
  

• 

  Western blot - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)

  All lanes : Anti-SMN/Gemin 1 antibody [2B1] (ab5831) at 1 µg/ml
  
  Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
  Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 35 kDa
  Observed band size: 37 kDawhy is the actual band size different from the predicted?
  Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.
  
  
  Exposure time: 20 minutes
  
  
  Gemin 1contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

• 

  Immunoprecipitation - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)

  Gemin 1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Gemin 1 (ab5831) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min,Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5831.
  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
  Band: 32kDa: Gemin 1

• 

  Flow Cytometry - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)

  Overlay histogram showing HepG2 cells stained with ab5831 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab5831, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 HepG2 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
  
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)Image courtesy of an anonymous Abreview.

  ab5831 staining Gemin 1 in human HeLa cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol and then blocked using 0.2% fish scale gelatin for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/300 for 20 minutes at 25°C. The secondary antibody used was a donkey anti-mouse IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. Counterstained with DAPI (blue).
  Gemin 1 is clearly visible in the cytoplasm and also as small dots in the nucleus (cajal bodies).

• 

  Western blot - Anti-SMN/Gemin 1 antibody [2B1] (ab5831)This image is courtesy of an abreview submitted by Samantha Jeschonek.

  Anti-SMN/Gemin 1 antibody [2B1] (ab5831) at 1/100 dilution + Xenopus laevis st II-III oocytes whole cell lysate at 20 µg
  
  Secondary
  Goat Anti-Mouse IgG Fc (HRP) (ab97265) at 1/25000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 35 kDa
  Observed band size: 31 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 30 seconds
  

  See Abreview