Anti-alpha smooth muscle Actin antibody [1A4]

• 产品名称

  Anti-alpha smooth muscle Actin抗体[1A4]
  参阅全部 alpha smooth muscle Actin 一抗

• 描述

  小鼠单克隆抗体[1A4] to alpha smooth muscle Actin

• 宿主

  Mouse

• 经测试应用

  适用于: ICC/IF, Flow Cyt (Intra), IHC-P, WBmore details

• 种属反应性

  与反应: Mouse, Rat, Human
  预测可用于: Sheep, Rabbit, Cow, Pig, Mammals, Baboon

• 免疫原

  Synthetic peptide corresponding to Human alpha smooth muscle Actin (N terminal).
  Database link: P62736

• 阳性对照

• WB: Human foreskin fibroblast lysate, human colon tissue lysate; NIH/3T3, SV40LT-SMC whole cell lysates. Flow Cyt (Intra): SV40LT-SMC. IHC-P: Human breast ductal carcinoma tissue. ICC/IF: SV40LT-SMC cell, SaOS-2, NIH3T3 WT and NIH3T3 KO cells

• 常规说明

  This antibody clone [1A4] 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. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.40
  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  1A4

• 同种型

  IgG2a

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab7817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: 101021287 Baboon

• Entrez Gene: 515610 Cow

• Entrez Gene: 59 Human

• Entrez Gene: 11475 Mouse

• Entrez Gene: 733615 Pig

• Entrez Gene: 100009271 Rabbit

• Entrez Gene: 81633 Rat

• Entrez Gene: 101121051 Sheep

• Omim: 102620 Human

• SwissProt: P62739 Cow

• SwissProt: P62736 Human

• SwissProt: P62737 Mouse

• SwissProt: P62740 Rabbit

• SwissProt: P62738 Rat

• Unigene: 500483 Human

• Unigene: 213025 Mouse

• Unigene: 195319 Rat

• Unigene: 3114 Rat

别名

• a actin antibody

• AAT6 antibody

• ACTA_HUMAN antibody

• ACTA2 antibody

• Actin alpha 2 smooth muscle aorta antibody

• Actin aortic smooth muscle antibody

• Actin, aortic smooth muscle antibody

• ACTSA antibody

• ACTVS antibody

• Alpha 2 actin antibody

• Alpha actin 2 antibody

• Alpha cardiac actin antibody

• alpha sma antibody

• Alpha-actin-2 antibody

• Cell growth inhibiting gene 46 protein antibody

• Cell growth-inhibiting gene 46 protein antibody

• GIG46 antibody

• Growth inhibiting gene 46 antibody

• MYMY5 antibody

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling alpha smooth muscle actin with ab7817 at a concentration of 0.01µg/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). ab7817 anti-alpha smooth muscle actin antibody [1A4] 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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an Abreview submitted by David Ivancic

  Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of 10% NBF-fixed human colon tissue permeabilized with 0.05% tween 20. Stained with ab7817 at 1/100 dilution. Secondary antibody used was Alexa fluor® 488 Donkey anti-Rabbit IgG at 1/300 dilution. Blocking was done with Sea Block for 30 minutes at 22°C. The sample was incubated with the primary antibody and Sea Block for 14 hours at 4°C. Antigen retrieval method was heat mediated, ab94674 100X Citrate Buffer pH 6.0.

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  ab7817 staining alpha smooth muscle Actin in SV40LT-SMC cells (positive control, top panel) and A431 cells (negative control, bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% 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 with ab7817 at 1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• 

  Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1 µg/ml
  
  Lane 1 : NIH 3T3 whole cell lysate
  Lane 2 : SV40LT-SMC whole cell lysate
  Lane 3 : A431 whole cell lysate
  Lane 4 : A549 whole cell lysate
  Lane 5 : Jurkat whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse IgG H&L (IRDye® 800RD) at 1/10000 dilution
  
  Observed band size: 42 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Gel type: MOPS

  Blocking buffer: 3% milk

  Loading control: alpha tubulin (ab52866), secondary Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (1:10000 dilution)

• 

  Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1 µg/ml
  
  Lane 1 : Human colon tissue lysate
  Lane 2 : Mouse colon tissue lysate
  Lane 3 : Human Foreskin Fibroblast Whole Cell Lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse IgG H&L (IRDye® 800RD) at 1/10000 dilution
  
  Observed band size: 42 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Gel type: MOPS

  Blocking buffer: 3% milk

  Loading control: alpha tubulin (ab52866), secondary Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (1:10000 dilution)

• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  This data was developed using the same antibody clone in a different buffer formulation that is PBS and sodium azide free (ab240654)

  ab240654 staining alpha smooth muscle Actin in SV40LT-SMC cells. The cells were fixed with 4% paraformaldehyde (10 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 ab240654 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 red). 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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  IHC image of alpha smooth muscle actin staining in a human breast ductal carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ 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 ab7817, 0.034µg/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.

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

• 

  Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)

  Overlay histogram showing SV40LT-SMC cells stained with ab7817 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab7817, 1.137µg/ml) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) at 1/2000 dilution for 30 min at 22°C

  Isotype control antibody (black line) was mouse IgG2a [18C8BC7AD10] (ab170191) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

  This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an anonymous Abreview

  Ab7817 staining alpha smooth muscle actin in Mouse intestine tissue by Immunohistochemistry-Immunofluorescence. Tissue was fixed with formaldehyde and blocked with 100% Cas-block for 30 minutes at room temperature; antigen retrieval was performed by heat mediated citrate buffer, pH6. The sample was incubated with primary antibody at 0.034µg/ml for 16 hours at 4°C. An Alexa Fluor® 488 Goat anti-mouse IgG was used as the secondary antibody at 1/400 dilution. Autofluorescence was blocked with 0.1% Sudan Black in 70% ethanol for 10 minutes at room temperature after antigen retrieval, and followed with 3X wash with PBS-T after antigen retrieval. Image was taken with confocal microscope.

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an Abreview submitted by Rudolf Jung

  ab7817 staining alpha smooth muscle actin in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized with wash buffer with tween; antigen retrieval was by heat mediation in Tris-EDTA buffer, pH 9.0. Samples were incubated with primary antibody (0.034µg/ml in blocking buffer) for 30 minutes at 20°C. A HRP-conjugated Goat anti-mouse IgG polyclonal (undiluted) was used as the secondary antibody.

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an anonymous Abreview

  Immunohistochemical analysis of mouse aorta (A) or skin (B) tissue, staining alpha smooth muscle Actin with ab7817.
  
  Tissue was fixed with 10% Neutral Buffered Formalin and blocked with 1% serum for 45 minutes 21°C; antigen retrieval was by enzymatic method in 0.0001% Trypsin-CaCl. Samples were incubated with primary antibody (0.034µg/ml in 0.3% Triton X-100 in PBS) for 1 hour at 21&degC. A biotin-conjugated horse anti-mouse polyclonal IgG (1/50) was used as the secondary antibody.

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an Abreview submitted by Charles Pallangyo

  ab7817 staining alpha smooth muscle Actin (green) in Mouse primary colon myofibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 5% BSA for 30 hours at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 5% BSA) for 2 hours at 25°C. Donkey Anti-Mouse IgG H&L (DyLight® 488) (ab96875) (1/1000) was used as the secondary antibody. Costained with ab92547, Rabbit anti-Vimentin (red).

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an anonymous abreview.

  ab7817 staining alpha smooth muscle Actin in human IMR-90 (Human Lung Fibroblast Cell Line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% TritonX-100 and blocked with 100% Cad-Block for 30 minutes at room temperature. Samples were incubated with primary antibody 3.41µg/ml in antibody diluent buffer for 16 hours at 4°C. An Alexa Fluor® 488-conjugated polyclonal Goat anti-mouse IgG, dilution 1/400, was used as secondary antibody.

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)This image is courtesy of an Abreview submitted by Dr. Ho-Jae Lee

  ab7817 staining alpha smooth muscle Actin in mouse heart cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with TritonX-100 and blocked with 5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody 6.82µg/ml in blocking buffer for 2 hours. An Alexa Fluor® 488-conjugated Donkey monoclonal to mouse IgG, dilution 1/200, was used as secondary antibody.

  See Abreview