Anti-GAPDH antibody [mAbcam 9484] - Loading Control

• 产品名称

  Anti-GAPDH抗体[mAbcam 9484] - Loading Control
  参阅全部 GAPDH 一抗

• 描述

  小鼠单克隆抗体[mAbcam 9484] to GAPDH - Loading Control

• 宿主

  Mouse

• 特异性

  In western blot, this product typically gives a lower signal in rat lysates compared to human and mouse lysates.

• 经测试应用

  适用于: WB, IHC-Pmore details

• 种属反应性

  与反应: Mouse, Rat, Chicken, Cow, Human, Pig, Xenopus laevis, Chinese hamster
  预测可用于: Rabbit, Dog, Cynomolgus monkey

• 免疫原

  Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB: A431, Jurkat, HEK293, HeLa, NIH 3T3, PC-12, EBTr and CHO cell lysates; Rat brain, chicken and pig liver and Xenopus laevis embryo tissue lysates. IHC-P: Human liver tissue.

• 常规说明

  For Western blotting, do not use milk for blocking. Our labs have extensively tested the blocking conditions for this antibody and recommend using 5% BSA for 1 hour. The comparison data is shown in the images section. 

  This antibody clone [mAbcam 9484] is manufactured by Abcam.

  If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

  
  

  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: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 浓度

• 10 µg 浓度为 1 mg/ml

• 100 µg 浓度为 1 mg/ml

• 纯度

  IgG fraction

• 克隆

  单克隆

• 克隆编号

  mAbcam 9484

• 骨髓瘤

  Sp2/0-Ag14

• 同种型

  IgG2b

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab9484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: 374193 Chicken

• Entrez Gene: 281181 Cow

• Entrez Gene: 403755 Dog

• Entrez Gene: 2597 Human

• Entrez Gene: 100042025 Mouse

• Entrez Gene: 14433 Mouse

• Entrez Gene: 396823 Pig

• Entrez Gene: 100009074 Rabbit

• Entrez Gene: 24383 Rat

• Entrez Gene: 685186 Rat

• Entrez Gene: 380259 Xenopus laevis

• Omim: 138400 Human

• SwissProt: P00356 Chicken

• SwissProt: P10096 Cow

• SwissProt: Q28259 Dog

• SwissProt: P04406 Human

• SwissProt: P16858 Mouse

• SwissProt: P00355 Pig

• SwissProt: P46406 Rabbit

• SwissProt: P04797 Rat

• SwissProt: P51469 Xenopus laevis

• 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

别名

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• 

  Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

  All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1 µg/ml
  
  Lane 1 : HeLa Whole Cell Lysate
  Lane 2 : NIH 3T3 Whole Cell Lysate
  Lane 3 : PC12 Whole Cell Lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 36 kDa
  
  
  Exposure time: 1 minute
  
  
  

  This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab9484 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

  IHC image of GAPDH staining in human liver FFPE section, performed on a 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 ab9484, 5µg/ml, for 8 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.

• 

  Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

  All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilution
  
  Lane 1 : Hela whole cell (Human)
  Lane 2 : 3T3 cell (Mouse)
  Lane 3 : Rat brain
  Lane 4 : Xenopus laevis embryo
  Lane 5 : Chicken Liver
  Lane 6 : EBTr cell (Cow)
  Lane 7 : CHO cell (Chinese hamster)
  Lane 8 : Pig liver
  
  Secondary
  All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 36 kDa
  Observed band size: 40 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 10 seconds
  
  
  

  The membrane was blocked in 5% BSA in TBST for 1 hour, then incubated for 1 hour in primary antibody diluted in TBST.

• 

  Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

  Lanes 1-5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% milk)
  Lanes 6-10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)
  
  Lanes 1 & 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
  Lanes 2 & 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lanes 3 & 8 : A-431 whole cell lysate (ab7909)
  Lanes 4 & 9 : Jurkat whole cell lysate (ab7899)
  Lanes 5 & 10 : HEK-293 whole cell lysate (ab7902)
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  Lanes 1-5 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
  Lanes 6-10 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
  
  Predicted band size: 36 kDa
  Observed band size: 40 kDawhy is the actual band size different from the predicted?
  
  
  
  

  The membrane 1-5 was blocked in 5% milk (1 hour). The membrane 6-10 was blocked in 5% BSA (1 hour). Milk is not a suitable blocking agent and significantly decreases the signal on the membrane.

• 

  Western blot - Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

  Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysate
  
  Secondary
  Goat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under non-reducing conditions.
  
  Predicted band size: 36 kDa
  
  
  Exposure time: 30 seconds