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    products/primary-antibodies/alpha-actininactn1-antibody-ep2528y-ab81265.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Crosslinking
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

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  • SDS
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Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
  • Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Human ACTN1 (alpha Actinin) knockout HeLa cell line (ab265610)

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Overview

  • Product name

    Anti-alpha Actinin/ACTN1 antibody [EP2528Y]
    See all alpha Actinin/ACTN1 primary antibodies
  • Description

    Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human alpha Actinin/ACTN1 aa 750-850 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HAP1, A431, NIH 3T3, C2C12, C6 and A673 cell lysates and human skeletal muscle and human heart tissue lysates.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP2528Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Crosslinking
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins
    • Stem Cells
    • Lineage Markers
    • Mesoderm
    • Developmental Biology
    • Lineage specification
    • Mesoderm

Associated products

  • Alternative Versions

    • Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • NIH/3T3 whole cell lysate (ab7179)
    • C2C12 whole cell lysate (ab7182)
  • Recombinant Protein

    • Recombinant Human alpha Actinin/ACTN1 protein (ab207132)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab81265 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
WB
1/500 - 1/1000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
Notes
WB
1/500 - 1/1000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein.
  • Sequence similarities

    Belongs to the alpha-actinin family.
    Contains 1 actin-binding domain.
    Contains 2 CH (calponin-homology) domains.
    Contains 2 EF-hand domains.
    Contains 4 spectrin repeats.
  • Cellular localization

    Cytoplasm > cytoskeleton. Cytoplasm > myofibril > sarcomere > Z line. Colocalizes with MYOZ2 and PPP3CA at the Z-line of heart and skeletal muscle.
  • Target information above from: UniProt accession P12814 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 87 Human
    • Entrez Gene: 109711 Mouse
    • Entrez Gene: 81634 Rat
    • Omim: 102575 Human
    • SwissProt: P12814 Human
    • SwissProt: Q7TPR4 Mouse
    • SwissProt: Q9Z1P2 Rat
    • Unigene: 509765 Human
    • Unigene: 253564 Mouse
    • Unigene: 403477 Mouse
    • Unigene: 6401 Rat
    see all
  • Alternative names

    • actinin 1 smooth muscle antibody
    • Actinin alpha 1 antibody
    • actinin, alpha 1 antibody
    • ACTN 1 antibody
    • Actn1 antibody
    • ACTN1_HUMAN antibody
    • Alpha Actinin 1 antibody
    • Alpha actinin cytoskeletal isoform antibody
    • Alpha-actinin cytoskeletal isoform antibody
    • Alpha-actinin-1 antibody
    • BDPLT15 antibody
    • F actin cross linking protein antibody
    • F-actin cross-linking protein antibody
    • FLJ40884 antibody
    • FLJ54432 antibody
    • Non muscle alpha actinin 1 antibody
    • Non-muscle alpha-actinin-1 antibody
    see all

Images

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : ACTN1 knockout HAP1 cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 103 kDa
    Observed band size: 115 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab81265 observed at 115 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab81265 was shown to react with ACTN1 in wild-type HAP1 cells in western blot with loss of signal observed in ACTN1 knockout sample. Wild-type and ACTN1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab81265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Alpha Actinin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: NIH 3T3 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab81265 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab81265 was shown to recognize alpha Actinin in wild-type HAP1 cells as signal was lost at the expected MW in alpha Actinin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab81265 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified)

    Lane 1 : NIH/3T3 whole cell lysate
    Lane 2 : C2C12 whole cell lysate
    Lane 3 : C6 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 103 kDa
    Observed band size: 103 kDa



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified) + A673 whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 103 kDa
    Observed band size: 103 kDa



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (unpurified)

    Lane 1 : NIH 3T3 cell lysate
    Lane 2 : human skeletal muscle tissue lysate
    Lane 3 : human heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 103 kDa
    Observed band size: 103 kDa

  • Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Protocols

  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab81265? Please let us know so that we can cite the reference in this datasheet.

ab81265 has not yet been referenced specifically in any publications.

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