- Tested Species Reactivity:
- Predicted Species Reactivity:
- Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Zebrafish
- Product Format:
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
- IHC, WB
- Reconstitution and Storage:
- For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
- Gene Symbol:
- Official Gene Full Name:
- Actinin, alpha 4
- NCBI Gene Id:
- Protein Name:
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Alias Symbols:
- DKFZp686K23158, FSGS, FSGS1, ACTININ-4
- Replacement Item:
- This antibody may replace item sc-134236 from Santa Cruz Biotechnology.
- Description of Target:
- Alpha actinins belong to the spectrin superfamily which represents a diverse group of cytoskeletal proteins, including the alpha and beta spectrins and dystrophins. Alpha actinin is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z-disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. ACTN4 is a nonmuscle, alpha actinin isoform which is concentrated in the cytoplasm, and thought to be involved in metastatic processes. Mutations in its gene have been associated with focal and segmental glomerulosclerosis.Alpha actinins belong to the spectrin gene superfamily which represents a diverse group of cytoskeletal proteins, including the alpha and beta spectrins and dystrophins. Alpha actinin is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z-disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. This gene encodes a nonmuscle, alpha actinin isoform which is concentrated in the cytoplasm, and thought to be involved in metastatic processes. Mutations in this gene have been associated with focal and segmental glomerulosclerosis.
- Protein Size (# AA):
- Molecular Weight:
- Protein A purified
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express ACTN4.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express ACTN4.
- The immunogen is a synthetic peptide directed towards the N terminal region of human ACTN4
- Predicted Homology Based on Immunogen Sequence:
- Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Zebrafish: 93%
- Complete computational species homology data:
- Anti-ACTN4 (ARP42202_T100)
- Peptide Sequence:
- Synthetic peptide located within the following region: LIHRHRPELIEYDKLRKDDPVTNLNNAFEVAEKYLDIPKMLDAEDIVNTA
- Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
- Protein Interactions:
- MYOZ2; HUWE1; UBC; BMP7; ACTN3; SUMO2; SUMO3; GNB2; SUMO1; NEDD8; MDM2; HGS; SF3A3; VCP; CDKN2A; ACTN1; PDLIM1; UBD; IGSF8; ICAM1; CD81; SVIL; Fcho2; POT1; VCAM1; TXN; RPS27; MAPK7; KPNA2; ITGA4; FN1; ESR1; CDH1; TRAF3IP1; FMNL1; NOS3; ACTN2; UBASH3B; SHC
- Blocking Peptide:
- For anti-ACTN4 (ARP42202_T100) antibody is Catalog # AAP42202 (Previous Catalog # AAPP24625)
- Printable datasheet for anti-ACTN4 (ARP42202_T100) antibody
- Sample Type Confirmation:
ACTN4 is supported by BioGPS gene expression data to be expressed in HCT116
- Target Reference:
- Kimura,K., (2006) Genome Res. 16 (1), 55-65
Wei, S., Gao, X., Du, J., Su, J. & Xu, Z. Angiogenin enhances cell migration by regulating stress fiber assembly and focal adhesion dynamics. PLoS One 6, e28797 (2011). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Zebrafish 22194915
Researcher: Johannes W. Hell, University of California
Application: Western blotting
Species + Tissue/Cell type: Lane1: 10 ug ACTN1-GFP transfected COS-7 lysate, Lane2: 10 ug ACTN2-GFP transfected COS-7 lysate, Lane3: 10 ug ACTN3-GFP transfected COS-7 lysate, Lane4: 10 ug ACTN4-GFP transfected COS-7 lysate
Primary antibody dilution: 1:!000
Secondary antibody: Anti-rabbit HRP
Secondary antibody dilution: 1:5000
How do Aviva's reagents play a role in your experimental goals?
Use in IF localization of a-actinin isoforms
How would you rate this antibody on a scale from 1-5 (5=best) and why?
5 specific signal in IF with overexpressed a-actinin-4
Would you use this antibody in future experiments?
Have you used another antibody which has worked in your application?
Yes, not isoform specific though
Do you believe the information about the reagent on Aviva's website is correct?
If the antibody works, do you plan to use it in future experiments or to publish your data?
How did you store the antibody after re-suspension?
Sample Description (please include species type and tissue/cell type):
Ectopic expression of all 4 a-actinin isiforms with GFP tag in COS-7 cells (monkey kidney fibroblast)
Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)?
4% paraformaldheyde/4%sucrose 5 minutes
How many different experimental trials were conducted using the antibody sample?
Primary antibody dilution, incubation time and temperature:
1:1000, O/N, 4C
Secondary antibody used, dilution, incubation time and temperature:
Alexa 568 anti-Rabbit, 2h, RT
From your IHC/ICC images, briefly explain the colors of each stain and counterstain:
Green: transfected GFP-tagged ACTN, Red Ab
Did you use an antigen retrieval method? If so, please explain?
What controls were used in your experiment?
Empty vector expressing GFP, other a-actinin isoforms
Please include your detailed tissue preparation and staining procedure/protocol here:
cDNA coding for the α-actinin isoforms 4 was subcloned into pEGFP vectors (Hall et al. unpublished). COS-7 cells were seeded onto glass coverslips and the next day transfected by calcium-phosphate precipitationwith the α-actinin constructs and empty pEGFP. After two days of expression, the cells were fixed (4% PFA/4% sucrose, 5 min at room temperature(RT)), permeabilized (0.25% Triton-X-100 in PBS, 10 min, RT), blocked (in PBS: 2% glycerol, 50 mM NH4Cl, 5% FBS, 2% normal goat serum, 2h, RT), and incubated with primary antibody (1:1000 in blocking buffer, overnight, 4C). Cells were blocked again (1h, RT), incubated with corresponding Alexa568-tagged secondary antibodies (1:100 in blocking buffer, 2h, RT), and the coverslips mounted on microscope slides (ProLong Gold antifade). All washing steps during staining were with PBS. Stained cells were visualized using an epifluorescence microscope with fluorescence filters for EGFP and TRITC.