製品の概要

  • 製品名
    Anti-beta Actin antibody [AC-15]
    beta Actin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [AC-15] to beta Actin
  • 特異性
    This antibody makes a good loading control antibody and we recommend its use as such. We also recommend ab8226, another excellent mouse monoclonal beta actin loading control antibody.
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, ICC, IHC-P, IHC-Fr, Indirect ELISA, WB, ELISAmore details
  • 種交差性
    交差種: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Human, Carp, African green monkey, Opossum, Chinese hamster, Common marmoset, Meriones unguiculatus
    非交差種: 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

  • エピトープ
    N-terminal of the beta isoform of actin.
  • ポジティブ・コントロール
    • Cultured human or chicken fibroblasts as described in Liao et al. ICC/IF: SV40LT-SMC cells
  • 特記事項

    Immunofluoresence staining of chicken gizzard ultra-thin sections (North et al. J. Cell Sci. 107:445-455 (1994)) labels the dense bodies, longitudinal channels and membrane associated dense-plaque.

    This product was changed from ascites to tissue culture supernatant on 31st January 2017. The following lots are from ascites and are still in stock on 31st January 2017- GR231981, GR247612 and GR181659. Lot numbers higher than GR247612 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 5 µg/ml.
IHC-FoFr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Indirect ELISA Use at an assay dependent concentration.
WB 1/5000 - 1/16000. Predicted molecular weight: 42 kDa.
ELISA Use at an assay dependent concentration.

ターゲット情報

画像



  • Predicted band size : 42 kDa

    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.

  • 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
    Goat Anti-Mouse IgG-Peroxidase
    Developed using the ECL technique

    Predicted band size : 42 kDa
  • 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).

  • Immunohistochemical frozen analysis of acetone-fixed human stomach tissue labeling beta Actin with ab6276 at 1/5000 dilution,followed by secondary antibody.

    See Abreview

  • 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
    Alexa Fluor anti mouse at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 42 kDa


  • Predicted band size : 42 kDa

    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.

    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 asssay. 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.



  • Predicted band size : 42 kDa
    Observed band size : 42 kDa

    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

参考文献

This product has been referenced in:
  • Li C  et al. CD54-NOTCH1 axis controls tumor initiation and cancer stem cell functions in human prostate cancer. Theranostics 7:67-80 (2017). Read more (PubMed: 28042317) »
  • Bademosi AT  et al. In vivo single-molecule imaging of syntaxin1A reveals polyphosphoinositide- and activity-dependent trapping in presynaptic nanoclusters. Nat Commun 8:13660 (2017). WB ; Drosophila melanogaster . Read more (PubMed: 28045048) »

See all 1015 Publications for this product

レビューと Q&A

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
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Abcam user community

Verified customer

投稿 Aug 01 2014

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
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Abcam user community

Verified customer

投稿 May 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Herr Dr. Vladimir Milenkovic

Verified customer

投稿 Jul 10 2013

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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Abcam user community

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投稿 Dec 12 2016

Abcam has not validated the combination of species/application used in this Abreview.
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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Abcam user community

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投稿 Nov 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
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
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Abcam user community

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投稿 Sep 23 2016

Abcam has not validated the combination of species/application used in this Abreview.
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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投稿 Jun 28 2016

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 Jun 19 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (spinal cord dorsal horn)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
30 µg
Specification
spinal cord dorsal horn
Blocking step
1x Rapid block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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投稿 Mar 03 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Human Foreskin Fibroblasts)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount
100000 cells
Specification
Human Foreskin Fibroblasts
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
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投稿 Jan 20 2016

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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