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Isotype/Loading Controls Loading Controls Beta Actin
使用敲除细胞株进行验证

Anti-beta Actin抗体[AC-15] (ab6276)

  • Datasheet
  • SDS
Reviews (94)Q&A (47)References (1672)

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Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [AC-15] (ab6276)
  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)

Key features and details

  • Mouse monoclonal [AC-15] to beta Actin
  • Suitable for: ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Hamster, Cow, Dog, African green monkey, Chinese hamster
  • Isotype: IgG1

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概述

  • 产品名称

    Anti-beta Actin抗体[AC-15]
    参阅全部 beta Actin 一抗
  • 描述

    小鼠单克隆抗体[AC-15] to beta Actin
  • 宿主

    Mouse
  • 经测试应用

    适用于: ICC/IF, WBmore details
  • 种属反应性

    与反应: Mouse, Rat, Hamster, Cow, Dog, African green monkey, Chinese hamster
    不与反应: Drosophila melanogaster, Dictyostelium discoideum
  • 免疫原

    Synthetic peptide corresponding to beta Actin aa 1-14 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). Slightly modified ß-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Al?a-Leu-Val-Ile-Asp-Asn-Gl y?-Ser-Gly-Lys, conjugated to KLH.
    Sequence:

    DDDIAALVIDNGSGK

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 表位

    N-terminal of the beta isoform of actin.
  • 阳性对照

    • ICC/IF: SV40LT-SMC cells WB: HAP1, HeLa, Jurkat, A431, HEK-293, COS-7, NIH/3T3, PC-12, Rat2, CHO, MDBK and MDCK cell lysates. IHC-Fr: Human stomach tissue. ICC/IF: SV40LT-SMC cells.
  • 常规说明

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

性能

  • 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 long term. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度

    Affinity purified
  • 纯化说明

    Purified from hybridoma cell culture.
  • 克隆

    单克隆
  • 克隆编号

    AC-15
  • 同种型

    IgG1
  • 研究领域

    • Isotype/Loading Controls
    • Loading Controls
    • Beta Actin
    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Actin
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin
    • Signal Transduction
    • Antibodies
    • beta actin

相关产品

  • Alternative Versions

    • Anti-Actin antibody [mAbGEa] (ab230169)
    • HRP Anti-beta Actin antibody [AC-15] (ab49900)
    • FITC Anti-beta Actin antibody [AC-15] (ab6277)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • Jurkat whole cell lysate (ab7899)
    • A431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human beta Actin protein (His tag) (ab240844)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Human beta Actin peptide (ab13772)
    • Phalloidin-iFluor 488 Reagent (ab176753)
    • Phalloidin-iFluor 647 Reagent (ab176759)
    • HRP Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab197277)
    • HRP Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab20272)
    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
    • Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
    • Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

应用

Our Abpromise guarantee covers the use of ab6276 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 5 µg/ml.
WB 1/5000 - 1/16000. Predicted molecular weight: 42 kDa.

靶标

  • 功能

    Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • 疾病相关

    Defects in ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYTJ patients manifest progressive, generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing loss.
  • 序列相似性

    Belongs to the actin family.
  • 翻译后修饰

    ISGylated.
  • 细胞定位

    Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Target information above from: UniProt accession P60709 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 280979 Cow
    • Entrez Gene: 403580 Dog
    • Entrez Gene: 11461 Mouse
    • Entrez Gene: 81822 Rat
    • SwissProt: P48975 Chinese hamster
    • SwissProt: P60712 Cow
    • SwissProt: O18840 Dog
    • SwissProt: P60710 Mouse
    • SwissProt: P60711 Rat
    • Unigene: 328431 Mouse
    • Unigene: 391967 Mouse
    • Unigene: 94978 Rat
    see all
  • 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
    see all

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [AC-15] (ab6276)
    Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [AC-15] (ab6276)

    ab6276 staining beta Actin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 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 1h. The cells were then incubated with ab6276 at a working concentration of 5μg/ml and ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    Western blot - Anti-beta Actin antibody [AC-15] (ab6276)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Beta actin knockout HAP1 cell lysate (20 µg)

    Lanes 1 and 2: Merged signal (red and green). Green - beta actin, ab6276 observed at 42 kDa. Red - loading control, ab181602 observed at 37 kDa.

    Ab6276 was shown to specifically react with beta actin in wild-type HAP1 cells. No band was observed when beta actin knockout samples were used. Wild-type and beta actin knockout samples were subjected to SDS-PAGE. ab6276 (beta actin) and ab181602 (loading control to GAPDH) were diluted 1/5000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    Western blot - Anti-beta Actin antibody [AC-15] (ab6276)This image is courtesy of an Abreview submitted by Dr Mark Elliott

    Western Blot of ab6276 (used as loading control) with whole tissue lysate of grey matter from BA20 (temporal cortex).  Ab6276 was diluted 1/50000 and incubated with the sample for 16 hours at 4°C.  5 µg of lysate was loaded onto the gel, which was blocked with 5% milk for 1 hour at 20°C.  An Alexa Fluor® 680 conjugated goat anti-mouse antibody, diluted 1/5000, was used as the secondary.

    Bands below actin in image are synaptophysin (SYN).

    See Abreview

  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1 µg/ml

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 3 : COS-7 (african green monkey kidney fibroblast-like cell line) cell lysate
    Lane 4 : NIH/3T3 (mouse embryonic fibroblast cell line) cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate
    Lane 6 : Rat2 (rat fibroblast cell line) cell lysate
    Lane 7 : CHO (chinese hamster ovary cell line) cell lysate
    Lane 8 : MDBK (bovine kidney cell line) cell lysate
    Lane 9 : MDCK (canine kidney cell line) cell lysate

    Secondary
    All lanes : Goat Anti-Mouse IgG-Peroxidase

    Developed using the ECL technique.

    Predicted band size: 42 kDa

  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1/5000 dilution

    Lane 1 : HeLa nuclear
    Lane 2 : HeLa whole cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : HEK293 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: 42 kDa
    Observed band size: 42 kDa

  • Western blot - Anti-beta Actin antibody [AC-15] (ab6276)
    Western blot - Anti-beta Actin antibody [AC-15] (ab6276)This image is courtesy of David Grimm, Yale University, USA.

    MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard assay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.

实验方案

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (1672)

    发表研究结果有使用 ab6276?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab6276 被引用在 1672 文献中.

    • Benfield CT  et al. Bat IFITM3 restriction depends on S-palmitoylation and a polymorphic site within the CD225 domain. Life Sci Alliance 3:N/A (2020). PubMed: 31826928
    • Dong Y  et al. Schizandrin A Protects Human Retinal Pigment Epithelial Cell Line ARPE-19 against HG-Induced Cell Injury by Regulation of miR-145. Mol Ther Nucleic Acids 19:42-49 (2020). PubMed: 31794890
    • Pungsrinont T  et al. Senolytic compounds control a distinct fate of androgen receptor agonist- and antagonist-induced cellular senescent LNCaP prostate cancer cells. Cell Biosci 10:59 (2020). PubMed: 32351687
    • Han X  et al. Advanced glycation end products enhance macrophage polarization to the M1 phenotype via the HIF-1a/PDK4 pathway. Mol Cell Endocrinol 514:110878 (2020). PubMed: 32464167
    • Akdag M  et al. Proximal Biotinylation-Based Combinatory Approach for Isolating Integral Plasma Membrane Proteins. J Proteome Res N/A:N/A (2020). PubMed: 32500712
    View all Publications for this product

    客户评价及客户问答

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    1-10 of 141 Abreviews or Q&A

    Western blot abreview for Anti-beta Actin antibody [AC-15]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (12)
    Sample
    Human Tissue lysate - whole (CNS tissue)
    Specification
    CNS tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 01 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-beta Actin [AC-15] antibody

    Excellent
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    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
    Sample
    Human Cell (HeLa cells)
    Specification
    HeLa cells
    Permeabilization
    Yes - 0.2% Triton-X100
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 May 23 2014

    Western blot abreview for Anti-beta Actin [AC-15] antibody

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    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing (12.5%)
    Sample
    Mouse Cell lysate - whole cell (Mouse brain lysates)
    Specification
    Mouse brain lysates
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Herr Dr. Vladimir Milenkovic

    Verified customer

    提交于 Jul 10 2013

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse neural stem cells)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    25 µg
    Specification
    mouse neural stem cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dr. Robert Kupp

    Verified customer

    提交于 Oct 19 2020

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (T-cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris HCl)
    Loading amount
    10.6 µg
    Specification
    T-cells
    Blocking step
    Licor intercept TBS as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

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    提交于 Feb 20 2020

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Application
    Western blot
    Sample
    Chinese Hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    50000 cells
    Specification
    Chinese hamster ovary (CHO) cells
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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    提交于 Dec 12 2016

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Application
    Western blot
    Sample
    Rabbit Cell lysate - whole cell (Rabbit Reticulocyte)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    0.5 µg
    Specification
    Rabbit Reticulocyte
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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    提交于 Nov 21 2016

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Application
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    Sample
    Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    50000 cells
    Specification
    Canine Kidney Epithelial MDCK cells
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

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    提交于 Sep 23 2016

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Application
    Western blot
    Sample
    African Green Monkey Cell lysate - whole cell (VERO E6)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    75000 cells
    Specification
    VERO E6
    Blocking step
    No blocking step used for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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    提交于 Jun 28 2016

    Western blot abreview for Anti-beta Actin antibody [AC-15]

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    Application
    Western blot
    Sample
    Hamster Tissue lysate - whole (BHK-21 (Kidney Fibroblasts))
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    50000 cells
    Specification
    BHK-21 (Kidney Fibroblasts)
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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