Anti-beta Actin antibody [mAbcam 8224] - Loading Control

• 产品名称

  Anti-beta Actin抗体[mAbcam 8224] - Loading Control
  参阅全部 beta Actin 一抗

• 描述

  小鼠单克隆抗体[mAbcam 8224] to beta Actin - Loading Control

• 宿主

  Mouse

• 特异性

  Recognises a single band at 42kD representing beta 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: WB, IHC-P, ICC/IFmore details
  不适用于: Flow Cyt (Intra)

• 种属反应性

  与反应: Mouse, Rat, Human, Xenopus laevis, Drosophila melanogaster, Schizosaccharomyces pombe
  预测可用于: Rabbit, Chicken, Cow, Cat, Dog, Pig, Chinese hamster, Other species

• 免疫原

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

• 阳性对照

• WB: A431; HEK293; NIH3T3; PC12 whole cell lysates; Xenopus embryo lysate; Drosophila lysate; S. pombe lysate. ICC/IF: HeLa cells. IHC/P: Human colon (FFPE)

• 常规说明

  This monoclonal antibody to beta actin works well as a protein loading control in Western blot for a broad range of species including Xenopus, Drosophila and S. pombe.

  This antibody clone [mAbcam 8224] 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). 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

• 250 µg 浓度为 1 mg/ml

• 纯度

  Affinity purified

• Primary antibody说明

  This clone works well as a loading control for Xenopus, Drosophila, S. cerevisiae and S.pombe. We recommend using ab8224 instead of ab8226 for these species.

• 克隆

  单克隆

• 克隆编号

  mAbcam 8224

• 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ab8224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。

应用说明

Is unsuitable for Flow Cyt (Intra).

数据库链接

• Entrez Gene: 396526 Chicken

• Entrez Gene: 280979 Cow

• Entrez Gene: 403580 Dog

• Entrez Gene: 60 Human

• Entrez Gene: 11461 Mouse

• Entrez Gene: 414396 Pig

• Entrez Gene: 100009272 Rabbit

• Entrez Gene: 81822 Rat

• Entrez Gene: 398459 Xenopus laevis

• Omim: 102630 Human

• SwissProt: P60706 Chicken

• SwissProt: P48975 Chinese hamster

• SwissProt: P60712 Cow

• SwissProt: O18840 Dog

• SwissProt: P60709 Human

• SwissProt: P60710 Mouse

• SwissProt: Q6QAQ1 Pig

• SwissProt: P29751 Rabbit

• SwissProt: P60711 Rat

• SwissProt: O93400 Xenopus laevis

• Unigene: 520640 Human

• Unigene: 708120 Human

• Unigene: 727576 Human

• Unigene: 328431 Mouse

• Unigene: 391967 Mouse

• Unigene: 94978 Rat

• Unigene: 4138 Xenopus laevis

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

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
  
  Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
  Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
  Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
  Lane 5 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue
  
  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 ab8224 overnight at 4°C. Antibody binding was detected using a goat anti-mouse 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 8224] - Loading Control (ab8224)

  IHC image of ab8224 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 ab8224, 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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

  ab8224 staining beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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 ab8224 at 1µg/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 8224] - Loading Control (ab8224)

  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
  
  Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
  Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
  Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  Exposure time: 3 minutes
  
  
  

  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 ab8224 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

  IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8224, 3µg/ml overnight at +4°C. A goat anti-mouse HRP-conjugated secondary antibody (ab6789, 1/2000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

  The inset negative control image is secondary-only at 1/500 dilution.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

  Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) + Xenopus embryo lysate at 20 µg
  
  Secondary
  Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  
  
  
  

  ab8224 used on Xenopus embryo lysate (20 ug of lysate/lane).
  
  Secondary
  Rabbit polyclonal anti-mouse HRP was used as the secondary antibody (ab6728) and developed using the ECL technique.
  Performed under reducing conditions.
  
  Predicted band size : 42kD

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1 µg/ml
  
  Lane 1 : Drosophila lysate
  Lane 2 : S. pombe lysate
  Lane 3 : S. cerevisiae lysate (Actin 1 - please see note)
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  
  

  Note: although S. cerevisiae is not known to express beta Actin, Abcam believes that the band on lane 3 corresponds to Actin 1 (Swissprot ID: P60010, based on sequence similarity).

   

  Secondary antibody - rabbit anti-mouse HRP (ab6728)

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)This image is courtesy of an anonymous Abreview

  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1/1000 dilution
  
  Lane 1 : Fruit fly (Drosophila melanogastor) whole cell lysate - Female
  Lane 2 : Fruit fly (Drosophila melanogastor) whole cell lysate -
  Male
  
  Lysates/proteins at 100 µg per lane.
  
  Secondary
  All lanes : An HRP-conjugated Sheep polyclonal to mouse IgG at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 42 kDa
  
  
  Exposure time: 2 minutes
  
  
  Blocking step: 5% Milk for 1 hour at 20°C.

  See Abreview

• 

  Western blot - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)This image is courtesy of an Abreview submitted by Joel Ohana

  Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1/10000 dilution + Mouse CT26 cells at 40 µg
  
  Secondary
  Anti-mouse IgG, HRP-linked Antibody at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  
  Exposure time: 6 minutes
  
  
  

  Western blot analysis using ab8224 at 1:1000 on Mouse CT26 cells. Blocking agent and dilution buffer was 5% milk in TBST

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

  Immunohistochemistical detection of beta Actin using antibody [mAbcam 8224] - Loading Control on formaldehyde-fixed paraffin-embedded rat cerebellum sections. Antigen retrieval step: heat mediated Citric acid pH6 buffer.  Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary antibody dilution 1/1000 for 2 hours in TBS/BSA/azide. Secondary Antibody: anti Mouse Igs conjugated to biotin (1/200). beta Actin appears to be particularly enriched not only in the glomeruli of the Granule cell layer (indicated by red arrowheads ) but also in Microglia (indicated by green arrowheads); All positive microglia appear to be ramified thus not presumed to be activated.

  See Abreview