Recombinant Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)
![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)](https://www.abcam.com/ps/products/239/ab239858/Images/ab239858-450837-anti-alpha-actinin-actn1-antibody-ep2527y-westernblot-hela-human.jpg "Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2527Y] to alpha Actinin/ACTN1 - BSA and Azide free
- Suitable for: ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free
See all alpha Actinin/ACTN1 primary antibodies -
Description
Rabbit monoclonal [EP2527Y] to alpha Actinin/ACTN1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC-12, C6, NIH/3T3, and HeLa cell lysates; IP: NIH/3T3 cells.
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General notes
ab239858 is the carrier-free version of ab68194.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2527Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239858 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 |
Use at an assay dependent concentration.
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| IP |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
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| Notes |
|---|
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ICC/IF
Use at an assay dependent concentration. |
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IP
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. 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 [EP2527Y] - BSA and Azide free (ab239858)Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/10000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 103 kDa
Observed band size: 103 kDaThis data was developed using ab68194, the same antibody clone in a different buffer formulation.
Blocking buffer: 5% NFDM/TBST
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![Immunoprecipitation - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)](https://www.abcam.com/ps/products/239/ab239858/Images/ab239858-4-anti-alpha-actinin-actn1-antibody-ep2527y-immunoprecipitation-nih-3t3-mouse.jpg)
Immunoprecipitation - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)Purified ab68194 at 1:20 dilution (2µg) immunoprecipitating alpha Actinin/ACTN1 in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10µg
Lane 2 (+): ab68194 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68194 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 103 kDa -
![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)](https://www.abcam.com/ps/products/239/ab239858/Images/ab239858-450836-anti-alpha-actinin-actn1-antibody-ep2527y-westernblot-mouse-rat.jpg)
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/1000 dilution (Purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 103 kDa
Observed band size: 103 kDaThis data was developed using ab68194, the same antibody clone in a different buffer formulation.
Blocking buffer: 5% NFDM/TBST
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![Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)](https://www.abcam.com/ps/products/239/ab239858/Images/ab239858-397589-anti-alpha-actininactn1-antibody-ep2527y-western-blot-hela-lysates.jpg)
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - BSA and Azide free (ab239858)All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/2000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ACTN1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 103 kDa
Observed band size: 103 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab68194).
Lanes 1- 2: Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab68194 was shown to react with alpha Actinin/ACTN1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265610 (knockout cell lysate ab257337) was used. Wild-type HeLa and ACTN1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab68194 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab239858 has not yet been referenced specifically in any publications.
