Alpha Actinin Monoclonal antibody

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  WB analysis using 66895-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66895-1-Ig (a-Actinin antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of RAW 264.7 using 66895-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 66895-1-Ig (a-Actinin antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse heart using 66895-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 66895-1-Ig (alpha Actinin antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse heart using 66895-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 66895-1-Ig (alpha Actinin antibody) at dilution of 1:1000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of C2C12 using 66895-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed C2C12 cells using Alpha Actinin antibody (66895-1-Ig, Clone: 1E2E1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  FC experiment of A431 using 66895-1-Ig

  1X10^6 A431 cells were intracellularly stained with 0.4 ug Anti-Human Alpha Actinin (66895-1-Ig, Clone:1E2E1) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Alpha Actinin Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 66895-1-Ig

Clone No.1E2E1

Publications(7)

Host / Isotype

Mouse / IgG2b

Reactivity

Human, rat, mouse and More (2)

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Synonyms

Alpha actinin 1, ACTN1, ACTN 1, actinin, alpha 1, actin

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Tested Applications

• Positive WB detected in: HeLa cells, RAW 264.7 cells, HEK-293 cells, HepG2 cells, Jurkat cells, HSC-T6 cells
• Positive IHC detected in: mouse heart tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: C2C12 cells
• Positive FC (Intra) detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 4 publications below
• IF: See 3 publications below

Product Information

66895-1-Ig targets Alpha Actinin in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with Human, rat, mouse samples.

• Tested Reactivity: Human, rat, mouse
• Cited Reactivity: human, mouse, rat, mosquito
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Alpha Actinin fusion protein Ag28383
• Full Name: actinin, alpha 1
• Calculated Molecular Weight: 103 kDa
• Observed Molecular Weight: 100-105 kDa
• GenBank Accession Number: BC015766
• Gene Symbol: Alpha Actinin
• Gene ID (NCBI): 87
• RRID: AB_2882224
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P12814
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Alpha Actinins are highly conserved actin-binding proteins that form antiparallel homodimers with the actin-binding domains on each end of the molecule, allowing crosslinking of actin molecules. There are four alpha actinin isoforms identified: two ubiquitously expressed cytoskeletal isoforms (ACTN1 and ACTN4) and two striated muscle-specific sarcomeric isoforms (ACTN2 and ACTN3). ACTN1 is also alternatively spliced to form smooth muscle or brain specific isoforms. This antibody recognize endogenous ACTN1, while it may cross-react with ACTN2 and ACTN4.

Publications

Species	Application	Title
rat	IF	Cell Mol Life Sci; SGK3 deficiency in macrophages suppresses angiotensin II-induced cardiac remodeling via regulating Ndufa13-mediated mitochondrial oxidative stress; Authors - Jiayu Ren
mosquito	WB	Cell Signal; Interplay between autophagy and Sindbis virus in cells derived from key arbovirus vectors, Aedes albopictus and Aedes aegypti mosquitoes; Authors - Jialu Qiao
human	IF	Front Pharmacol; Chronic Administration of COVID-19 Drugs Fluvoxamine and Lopinavir Shortens Action Potential Duration by Inhibiting the Human Ether-à-go-go-Related Gene and Cav1.2; Authors - Zequn Zheng
rat	IF	Cardiovasc Diabetol; Dysregulated Rbfox2 produces aberrant splicing of CaV1.2 calcium channel in diabetes-induced cardiac hypertrophy; Authors - Pengpeng Li
mouse	WB	Nucleic Acid Ther; Challenges of Assessing Exon 53 Skipping of the Human DMD Transcript with Locked Nucleic Acid-Modified Antisense Oligonucleotides in a Mouse Model for Duchenne Muscular Dystrophy; Authors - Sarah Engelbeen
human,mouse	WB	Cell Mol Biol Lett; APOE2 protects against Aβ pathology by improving neuronal mitochondrial function through ERRα signaling; Authors - Zhiyuan Ning