Anti-beta Actin antibody [mAbcam 8226] - Loading Control

• 产品名称

  Anti-beta Actin抗体[mAbcam 8226] - Loading Control
  参阅全部 beta Actin 一抗

• 描述

  小鼠单克隆抗体[mAbcam 8226] to beta Actin - Loading Control

• 宿主

  Mouse

• 特异性

  Does not cross-react with adult cardiac, smooth, or skeletal muscle actin. The immunogen used for this product shares 77% homology with Gamma actin/actin cytoplasmic 2. Cross-reactivity with this protein has not been confirmed experimentally.

• 经测试应用

  适用于: ICC/IF, IHC-P, WBmore details

• 种属反应性

  与反应: Mouse, Rat, Human
  预测可用于: Sheep, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Pig, Monkey, Zebrafish, African green monkey, Chinese hamster, Armenian hamster

• 免疫原

  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB, ICC/IF: HeLa, Jurkat, A431, HEK293, NIH 3T3, PC12 cells. IHC-P: Human colon (FFPE), Rat colon (FFPE) tissue. Hela cells

• 常规说明

  Western blot protocol advice:

  We recommend blocking with 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please see the comparison data in the images section. If milk block is required, we recommend using ab8224 mouse monoclonal [mAbcam 8224] to beta actin. Contact our Scientific Support team for more information or advice.

  This antibody clone [mAbcam 8226] 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

• 100 µg 浓度为 1 mg/ml

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  mAbcam 8226

• 骨髓瘤

  Sp2/0-Ag14

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab8226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: 396526 Chicken

• Entrez Gene: 280979 Cow

• Entrez Gene: 403580 Dog

• Entrez Gene: 100033878 Horse

• Entrez Gene: 60 Human

• Entrez Gene: 11461 Mouse

• Entrez Gene: 414396 Pig

• Entrez Gene: 100009272 Rabbit

• Entrez Gene: 81822 Rat

• Entrez Gene: 443052 Sheep

• Entrez Gene: 57934 Zebrafish

• Omim: 102630 Human

• SwissProt: P60706 Chicken

• SwissProt: P48975 Chinese hamster

• SwissProt: P60712 Cow

• SwissProt: O18840 Dog

• SwissProt: P60708 Horse

• SwissProt: P60709 Human

• SwissProt: P60710 Mouse

• SwissProt: Q6QAQ1 Pig

• SwissProt: P29751 Rabbit

• SwissProt: P60711 Rat

• SwissProt: Q7ZVI7 Zebrafish

• Unigene: 520640 Human

• Unigene: 708120 Human

• Unigene: 727576 Human

• Unigene: 328431 Mouse

• Unigene: 391967 Mouse

• Unigene: 94978 Rat

• Unigene: 35143 Zebrafish

别名

• A26C1A antibody

• A26C1B antibody

• ACTB antibody

• ACTB_HUMAN antibody

• Actin beta antibody

• Actin cytoplasmic 1 antibody

• Actin, cytoplasmic 1, N-terminally processed antibody

• Actx antibody

• b actin antibody

• b-actin antibody

• Beta cytoskeletal actin antibody

• Beta-actin antibody

• BRWS1 antibody

• E430023M04Rik antibody

• Melanoma X actin antibody

• MGC128179 antibody

• PS1TP5 binding protein 1 antibody

• PS1TP5BP1 antibody

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling beta actin with ab8226 at a concentration of 0.06µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab8226 anti-beta actin antibody [mAbcam 8226] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  This data was developed using the same antibody clone in a different buffer formulation containing only PBS (ab264083). ab264083 staining beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab264083 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
  
  Lane 1 : A431 (Human epidermoid carcinoma cell line) Whole Cell Lysate
  Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
  Lane 3 : NIH 3T3 (Mouse embryo fibroblast cell line) Whole Cell Lysate
  Lane 4 : PC12 (Rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at 1/10000 dilution
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  
  

  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 5% Milk before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  IHC image of ab8226 staining beta Actin in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. 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 ab8226, 0.1μg/ml working concentration, 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. No primary antibody was used in the secondary only control (shown on the inset).
  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.
  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  IHC image of ab8226 staining beta Actin in rat colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab8226, 0.5μg/ml working concentration, 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. No primary antibody was used in the secondary only control (shown on the inset).
  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-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  Lane 1 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
  Lane 2 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution
  Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution
  
  Lanes 1-2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
  Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate
  Lane 4 : NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  
  
  Exposure time: 10 seconds
  

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml
  
  Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
  Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
  Lane 4 : HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
  Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : HRP-conjugated goat anti-mouse IgG at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  Exposure time: 10 seconds
  
  
  

  Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) with the Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623).

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

  Lanes 1-3 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% BSA BLOCK)
  Lanes 4-6 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% MILK BLOCK)
  
  Lanes 1 & 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lanes 2 & 5 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
  Lanes 3 & 6 : NIH/3T3 (Mouse embryo fibroblast 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) at 1/3000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  Exposure time: 8 minutes