ACTN4 Polyclonal antibody

• Featured Product
• KD/KO Validated

ACTN4 Polyclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IP, IHC, IF, FC, CoIP, ELISA

Conjugate

Unconjugated

Publications

17

Cat no : 19096-1-AP

Synonyms

ACTININ 4, actinin, alpha 4, ACTN4, Alpha actinin 4, F actin cross linking protein, FSGS, FSGS1, Non muscle alpha actinin 4

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Validation Data Gallery

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  WB analysis using 19096-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 19096-1-AP (ACTN4 antibody) at dilution of 1:25000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse liver using 19096-1-AP

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 19096-1-AP (ACTN4 Antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 19096-1-AP

  IP result of anti-ACTN4 (IP:19096-1-AP, 5ug; Detection:19096-1-AP 1:1000) with HeLa cells lysate 3000ug.

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  IHC staining of human lung cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 19096-1-AP (ACTN4 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 19096-1-AP (ACTN4 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 19096-1-AP (ACTN4 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 19096-1-AP (ACTN4 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer using 19096-1-AP (ACTN4 antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human lung cancer using 19096-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer using 19096-1-AP (ACTN4 antibody) at dilution of 1:100 (under 40x lens).

• 

  IF Staining of MCF-7 using 19096-1-AP

  Immunofluorescent analysis of MCF-7 cells, using ACTN4 antibody 19096-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

• 

  IF Staining of HeLa using 19096-1-AP

  Immunofluorescent analysis of Hela cells, using ACTN4 antibody 19096-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

• 

  IF Staining of MCF-7 using 19096-1-AP

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using 19096-1-AP (ACTN4 antibody) at dilution of 1:50 and Alexa Fluor 488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of HeLa using 19096-1-AP

  1X10^6 HeLa cells were stained with 0.2ug ACTN4 antibody (19096-1-AP, red) and control antibody (blue). Fixed with 90% MeOH blocked with 3% BSA (30 min). Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) with dilution 1:1000.

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Tested Applications

• Positive WB detected in: HEK-293T cells, mouse liver tissue, HeLa cells, Jurkat cells, C2C12 cells, NIH/3T3 cells, mouse skeletal muscle, rat skeletal muscle
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung cancer tissue, human breast cancer 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 detected in: MCF-7 cells, Hela cells
• Positive FC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:10-1:100
• Flow Cytometry (FC): FC : 0.20 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

• KD/KO: See 2 publications below
• WB: See 13 publications below
• IHC: See 2 publications below
• IF: See 5 publications below
• ELISA: See 1 publications below
• CoIP: See 1 publications below

Product Information

19096-1-AP targets ACTN4 in WB, IP, IHC, IF, FC, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: ACTN4 fusion protein Ag13546
• Full Name: actinin, alpha 4
• Calculated Molecular Weight: 105 kDa
• Observed Molecular Weight: 100-105 kDa
• GenBank Accession Number: BC005033
• Gene Symbol: ACTN4
• Gene ID (NCBI): 81
• RRID: AB_10642150
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• 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

ACTN4 (alpha actinin 4) is a ubiquitous actin-binding protein that cross-links actin filaments into bundles to form filopodia, and has a crucial role in cytoskeletal organization. ACTN4 is colocalized with actin stress fibers and is dispersed in the cytoplasm and in the nucleus. Increased levels of ACTN4 have been associated with cancer metastasis, indicating its implication in cancer cell migration and metastasis. Defects in ACTN4 cause glomerular disease. This antibody recognizes 100-105 kDa of endogenous ACTN4 protein in various lysates. (22370640, 10700177)

Publications

Species	Application	Title
human	ELISA	J Thorac Oncol; Alpha-actinin 4 is Associated with Cancer Cell Motility and is a Potential Biomarker in Non-Small-Cell Lung Cancer.; Authors - Ming-Chuan Wang
human	WB	Oncogene; MTBP suppresses cell migration and filopodia formation by inhibiting ACTN4.; Authors - Agarwal N N
human	WB	Cancer Immunol Immunother; Circulating PD-L1 is associated with T cell infiltration and predicts prognosis in patients with CRLM following hepatic resection.; Authors - Xiuxing Chen
rat	WB,IF	Sci Rep; Poly-L-ornithine enhances migration of neural stem/progenitor cells via promoting α-Actinin 4 binding to actin filaments.; Authors - Hongfei Ge
mouse	IF	J Cell Sci; GIPC3 couples to MYO6 and PDZ domain proteins and shapes the hair cell apical region; Authors - Paroma Chatterjee
human	WB	Amino Acids; Quantitative proteomic dissection of a native 14-3-3ε interacting protein complex associated with hepatocellular carcinoma.; Authors - Chen Bai

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