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    products/primary-antibodies/beta-actin-antibody-ac-15-ab6276.pdf

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Isotype/Loading Controls Loading Controls Beta Actin
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Validated using a knockout cell line

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

  • Datasheet
  • SDS
Reviews (94)Q&A (47)References (1949)

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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, Cow, Dog, Human, African green monkey, Chinese hamster
  • Isotype: IgG1

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Overview

  • Product name

    Anti-beta Actin antibody [AC-15]
    See all beta Actin primary antibodies
  • Description

    Mouse monoclonal [AC-15] to beta Actin
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Dog, Human, African green monkey, Chinese hamster
    Does not react with: Drosophila melanogaster, Dictyostelium discoideum
  • Immunogen

    Synthetic peptide corresponding to beta Actin aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin. 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.

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

    N-terminal of the beta isoform of actin.
  • Positive control

    • 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.
  • General notes

    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

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

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

    Affinity purified
  • Purification notes

    Purified from hybridoma cell culture.
  • Clonality

    Monoclonal
  • Clone number

    AC-15
  • Isotype

    IgG1
  • Research areas

    • 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

Associated products

  • 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)
  • Isotype control

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

    • Jurkat whole cell lysate (ab7899)
    • A-431 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)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC/IF (9)
Use a concentration of 5 µg/ml.
WB (76)
1/5000 - 1/16000. Predicted molecular weight: 42 kDa.
Notes
ICC/IF
Use a concentration of 5 µg/ml.
WB
1/5000 - 1/16000. Predicted molecular weight: 42 kDa.

Target

  • Function

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

    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.
  • Sequence similarities

    Belongs to the actin family.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    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
  • Database links

    • Entrez Gene: 280979 Cow
    • Entrez Gene: 403580 Dog
    • Entrez Gene: 60 Human
    • Entrez Gene: 11461 Mouse
    • Entrez Gene: 81822 Rat
    • Omim: 102630 Human
    • SwissProt: P48975 Chinese hamster
    • SwissProt: P60712 Cow
    • SwissProt: O18840 Dog
    • SwissProt: P60709 Human
    • SwissProt: P60710 Mouse
    • SwissProt: P60711 Rat
    • Unigene: 520640 Human
    • Unigene: 708120 Human
    • Unigene: 727576 Human
    • Unigene: 328431 Mouse
    • Unigene: 391967 Mouse
    • Unigene: 94978 Rat
    see all
  • Alternative names

    • 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

Images

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

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

    Secondary
    All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution

    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.

  • 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 human 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.

Protocols

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1949)

Publishing research using ab6276? Please let us know so that we can cite the reference in this datasheet.

ab6276 has been referenced in 1949 publications.

  • Itai T  et al. De novo variants in CELF2 that disrupt the nuclear localization signal cause developmental and epileptic encephalopathy. Hum Mutat 42:66-76 (2021). PubMed: 33131106
  • Cooley MM  et al. Deficient Endoplasmic Reticulum Acetyl-CoA Import in Pancreatic Acinar Cells Leads to Chronic Pancreatitis. Cell Mol Gastroenterol Hepatol 11:725-738 (2021). PubMed: 33080365
  • Fløyel T  et al. SKAP2, a Candidate Gene for Type 1 Diabetes, Regulates ß-Cell Apoptosis and Glycemic Control in Newly Diagnosed Patients. Diabetes 70:464-476 (2021). PubMed: 33203694
  • Choi YS  et al. Chloroform extract of Citrus unshiu Markovich peel induces apoptosis and inhibits stemness in HeLa human cervical cancer cells. Mol Med Rep 23:N/A (2021). PubMed: 33236129
  • Fujihara K  et al. Genetic deletion of the 67-kDa isoform of glutamate decarboxylase alters conditioned fear behavior in rats. FEBS Open Bio 11:340-353 (2021). PubMed: 33325157
View all Publications for this product

Customer reviews and Q&As

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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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 01 2014

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

Excellent
Abreviews
Abreviews
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 23 2014

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

Excellent
Abreviews
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

HERR DR. Vladimir Milenkovic

Verified customer

Submitted Jul 10 2013

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

Good
Abreviews
Abreviews
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Robert Kupp

Verified customer

Submitted Oct 19 2020

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

Excellent
Abreviews
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 20 2020

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

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Dec 12 2016

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

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Nov 21 2016

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

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Sep 23 2016

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

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Jun 28 2016

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

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Jun 19 2016

1-10 of 141 Abreviews or Q&A

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