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
-
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.
-
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.
-
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.
Properties
-
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...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2527Y -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Recombinant Protein
-
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 |
---|---|---|
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).
|
Notes |
---|
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
-
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
-
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
-
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
-
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.
-
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
-
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.
-
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.
-
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
-
SDS download
-
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