Recombinant Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)
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
- 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]
See all alpha Actinin/ACTN1 primary antibodies -
Description
Rabbit monoclonal [EP2527Y] to alpha Actinin/ACTN1 -
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 corresponding to Human alpha Actinin/ACTN1.
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Positive control
- ICC/IF: ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612); IP: NIH/3T3 whole cell lysate; WB: HeLa, PC-12, NIH/3T3, and C6 whole cell 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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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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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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68194 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 |
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ICC/IF |
Use a concentration of 0.5 - 5 µg/ml.
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IP |
1/20.
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WB |
1/1000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
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Notes |
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ICC/IF
Use a concentration of 0.5 - 5 µg/ml. |
IP
1/20. |
WB
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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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 -
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 kDaBlocking buffer: 5% NFDM/TBST
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Immunofluorescence staining of Actinin/ACTN1 using ab68194 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 3 (left panel), 5 (middle panel) and 10 days (right panel) post induction.
The cells were fixed with 100% MeOH (5 min) and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68194 at 0.5 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.
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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 kDaBlocking buffer: 5% NFDM/TBST
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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 kDaLanes 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.
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Immunofluorescence staining of Actinin/ACTN1 using ab68194 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 3 (left panel), 5 (middle panel) and 10 days (right panel) post induction.
The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68194 at 0.5 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Alpha Actinin/ACTN1 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 - ab68194 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab68194 was shown to specifically react with alpha Actinin/ACTN1 in wild-type HAP1 cells as signal was lost in alpha Actinin/ACTN1 knockout cells. Wild-type and alpha Actinin/ACTN1 knockout samples were subjected to SDS-PAGE. ab68194 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab68194 has been referenced in 10 publications.
- Gajawada P et al. Myocardial Accumulations of Reg3A, Reg3? and Oncostatin M Are Associated with the Formation of Granulomata in Patients with Cardiac Sarcoidosis. Int J Mol Sci 22:N/A (2021). PubMed: 33923774
- Burton KM et al. Distinct forms of the actin cross-linking protein α-actinin support macropinosome internalization and trafficking. Mol Biol Cell 32:1393-1407 (2021). PubMed: 34010028
- Valussi M et al. Repression of Osmr and Fgfr1 by miR-1/133a prevents cardiomyocyte dedifferentiation and cell cycle entry in the adult heart. Sci Adv 7:eabi6648 (2021). PubMed: 34644107
- Gui M et al. Huoxue Qianyang Qutan recipe attenuates Ang II-induced cardiomyocyte hypertrophy by regulating reactive oxygen species production. Exp Ther Med 22:1446 (2021). PubMed: 34721688
- Liu Y et al. JMY expression by Sertoli cells contributes to mediating spermatogenesis in mice. FEBS J 287:5478-5497 (2020). PubMed: 32279424
- Burton KM et al. Dynamin 2 interacts with a-actinin 4 to drive tumor cell invasion. Mol Biol Cell 31:439-451 (2020). PubMed: 31967944
- Xie GF et al. High ACTN1 Is Associated with Poor Prognosis, and ACTN1 Silencing Suppresses Cell Proliferation and Metastasis in Oral Squamous Cell Carcinoma. Drug Des Devel Ther 14:1717-1727 (2020). PubMed: 32440097
- Kubin T et al. The MEK/ERK Module Is Reprogrammed in Remodeling Adult Cardiomyocytes. Int J Mol Sci 21:N/A (2020). PubMed: 32882982
- Jahan F et al. Phosphorylation of the α-chain in the integrin LFA-1 enables β2-chain phosphorylation and α-actinin binding required for cell adhesion. J Biol Chem 293:12318-12330 (2018). PubMed: 29903913
- Zong J et al. Nuclear Localization Leucine-Rich-Repeat Protein 1 Deficiency Protects Against Cardiac Hypertrophy by Pressure Overload. Cell Physiol Biochem 48:75-86 (2018). PubMed: 30001530