Recombinant Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-415036-anti-alpha-actininactn1-antibody-ep2528y-western-blot-hap1-wt-empty-hap1-actn1-knockout-a431-daudi.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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.
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Positive control
- WB: HAP1, A431, NIH 3T3, C2C12, C6 and A673 cell lysates and human skeletal muscle and human heart tissue lysates.
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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).
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| Notes |
|---|
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WB
1/500 - 1/1000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa). |
Target
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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. - Information by UniProt
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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
see all -
Alternative names
- actinin 1 smooth muscle antibody
- Actinin alpha 1 antibody
- actinin, alpha 1 antibody
see all
Images
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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.
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![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-299870-anti-alpha-actinin-antibody-ep2528y-western-blot.jpg)
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.
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![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-257344-anti-alpha-actinin-antibody-ep2528y-western-blot.jpg)
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)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-257345-anti-alpha-actinin-antibody-ep2528y-western-blot.jpg)
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)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-96445-anti-alpha-actinin-antibody-ep2528y-western-blot.jpg)
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)](https://www.abcam.com/ps/products/81/ab81265/Images/ab81265-7-benefits-of-recombinant-antibodies.png)
Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab81265 has not yet been referenced specifically in any publications.