Anti-GAPDH antibody [6C5] - Loading Control

• 产品名称

  Anti-GAPDH抗体[6C5] - Loading Control
  参阅全部 GAPDH 一抗

• 描述

  小鼠单克隆抗体[6C5] to GAPDH - Loading Control

• 宿主

  Mouse

• 特异性

  This GAPDH antibody can be used as a loading control antibody. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. There is no cross-reaction with GAPDH from yeast. Preliminary data indicates that the GAPDH antibody- loading control ab8245 recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.

• 经测试应用

  适用于: WB, ICC/IFmore details

• 种属反应性

  与反应: Mouse, Rat, Human
  预测可用于: Horse, Chicken, Guinea pig, Hamster, Cat, Dog, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, Xenopus tropicalis不与反应: Goat, Cow, Saccharomyces cerevisiae

• 免疫原

  Full length native protein (purified) corresponding to GAPDH.
  Database link: P46406

• 阳性对照

• ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells. WB: HeLa, A431, Jurkat, HEK-293, Raji whole cell lysate.

• 常规说明

  This product switched from ascites to tissue culture supernatant on 31 July 2017. Lot numbers higher than [GR291713] will be from tissue culture supernatant.

  
  

  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

  pH: 7.40
  Preservative: 0.09% Sodium azide
  Constituent: PBS

• 浓度

• 100 µg 浓度为 2 mg/ml

• 纯度

  Protein A purified

• 纯化说明

  Chromatography on protein A Sepharose

• 克隆

  单克隆

• 克隆编号

  6C5

• 骨髓瘤

  Sp2/0

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab8245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: 493876 Cat

• Entrez Gene: 374193 Chicken

• Entrez Gene: 403755 Dog

• Entrez Gene: 2597 Human

• Entrez Gene: 100042025 Mouse

• Entrez Gene: 14433 Mouse

• Entrez Gene: 396823 Pig

• Entrez Gene: 24383 Rat

• Entrez Gene: 685186 Rat

• Entrez Gene: 380259 Xenopus laevis

• Entrez Gene: 317743 Zebrafish

• Omim: 138400 Human

• SwissProt: Q9N2D5 Cat

• SwissProt: P00356 Chicken

• SwissProt: Q28259 Dog

• SwissProt: P04406 Human

• SwissProt: P16858 Mouse

• SwissProt: P00355 Pig

• SwissProt: P04797 Rat

• SwissProt: P51469 Xenopus laevis

• SwissProt: Q5XJ10 Zebrafish

• Unigene: 544577 Human

• Unigene: 592355 Human

• Unigene: 598320 Human

• Unigene: 304088 Mouse

• Unigene: 309092 Mouse

• Unigene: 317779 Mouse

• Unigene: 343110 Mouse

• Unigene: 392463 Mouse

• Unigene: 392480 Mouse

• Unigene: 414470 Mouse

• Unigene: 458138 Mouse

• Unigene: 458416 Mouse

• Unigene: 475698 Mouse

• Unigene: 129558 Rat

• Unigene: 91450 Rat

• Unigene: 995 Xenopus laevis

• Unigene: 35640 Zebrafish

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• 

  Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)This image is courtesy of an anonymous Abreview

  All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  
  Lane 1 : Mouse hippocampus whole cell lysate
  Lane 2 : Rat hippocampus whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : HRP-conjugated Rabbit anti-mouse at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 40.2 kDa
  Observed band size: 36 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 10 seconds
  

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

  ab8245 staining GAPDH in SV40LT-SMC (Rat SV40-transfected aorta smooth cell line) cells.

  The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

  Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 10 µg/ml + Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
  
  Predicted band size: 40.2 kDa
  
  

• 

  Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

  All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 2.5 µg/ml
  
  Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Nuclear
  Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
  Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
  Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Alexa Fluor anti-mouse at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 40.2 kDa
  Observed band size: 37 kDawhy is the actual band size different from the predicted?
  
  
  
  Fluorescence detection of secondary antibody.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

  ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

  The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)

  ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

  The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

  Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.