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

Anti-beta Actin antibody [AC-15] 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 - ab6276 observed at 42 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab6276 was shown to specifically react with beta actin when beta actin knockout samples were used. Wild-type and beta actin knockout samples were subjected to SDS-PAGE. ab6276 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
  • Paraffin-embedded human heart tissue stained for beta Actin using ab6276 at 10 µg/ml in immunohistochemical analysis.

  • 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
  • ab6276 at 1/500 staining human macrophage cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. A Cy2 ® conjugated donkey antibody was used as the secondary.

    See Abreview

  • Anti-beta Actin antibody [AC-15] (ab6276) at 1/20000 dilution + Hamster cell BHK-21 (Kidney Fibroblasts) lysate at 50000 cells

    Secondary
    Goat anti-mouse IgG H&L (IRDye® 800CW) at 1/15000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 43 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview.

    Detection: Near-Infrared Fluorescence Imaging.

     

    See Abreview



  • 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.

  • ab6276 staining beat actin in rat hypothalamus primary cells by ICC/IF. The cells were formaldehyde fixed, permeabilized in 0.5% Triton X-100 and blocked in 5% serum for 20 minutes at 25°C. The primary antibody was diluted 1/1000 in PBS (0.1% Triton X-100, 1% goat serum) and incubated with sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat polyclonal to mouse IgG1, diluted 1/500 was used as secondary.

    See Abreview

  • Anti-beta Actin antibody [AC-15] (ab6276) at 1/20000 dilution + Rabbit Reticulocyte whole cell lysate at 0.5 µg

    Secondary
    Goat anti-mouse IgG H&L (IRDye® 680RD) at 1/15000 dilution

    Predicted band size : 42 kDa
    Observed band size : 43 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous abreview.

    Reduced, denatured 4-12% Bolt Bis-Tris Plus gel. 
    Blocking: 100% Li-Cor Odyssey Blocking Buffer (TBS).

    See Abreview



  • 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

  • ab6276 at 1/10000 staining mouse brain tissue sections by IHC-P. The mouse brain was immersion-fixed in 4% Formalin/PBS for 2 days, then bisected mid-saggitally. The tissue was further fixed in same solution for a further 3 days. Both halves were processed to pwax on a dip-n-dunk tissue processor using a 20hr-cycle. Sections were cut at 5microns,floated out on water at 47°C, mounted on Superfrost Plus slides and dried/melted for 24hrs in a 62°C oven.

    The whole brain shows positivity but there are areas/cells that are enriched. Note that the axonal tracks are negatively stained at this dilution factor, yet some cells within those tracks are still strongly positive (many similar cells outside of the tracks are also positive).

    See Abreview

  • ab6276 detecting beta actin in AGS Human gastric carcinoma cells by direct ELISA. Samples were blocked with 5% BSA for 1 hour at 23°C and incubated with the primary antibody (1/1000 in blocking buffer) for 16 hours at 4°C. Ab6729 (1/1000) was used as the secondary antibody.

    See Abreview

Anti-beta Actin antibody [AC-15] (ab6276) 使用論文

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 962 Publications for this product

Product Wall

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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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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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
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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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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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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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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
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投稿 Jun 19 2016

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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"