Invitrogen

Phospho-Cofilin (Ser3) Polyclonal Antibody

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Phospho-Cofilin (Ser3) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Phospho-Cofilin pSer3 was done on 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Cofilin pSer3 Rabbit Polyclonal Antibody (Product # 44-1072G) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Phospho-Cofilin (Ser3) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (20 µg lysate) of HeLa (Lane 1), HeLa treated for overnight with 150 uM of H2O2 (Lane 2), HeLa treated for overnight with 3 uM of Staurosporine (lane 3), COS-7 (lane 4), NIH/3T3 (lane 5), MCF7 (lane 6), Rat Skeletal Muscle (lane 7), A-431 (lane 8), A-431 treated for overnight with 150 uM of H2O2 (lane 9), Jurkat (lane 10) and Jurkat treated for overnight with 3 uM of Staurosporine (lane 11). The blots were probed with Anti-Cofilin (pS3) Rabbit Polyclonal Antibody (Product # 44-1072G, 1-3 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 18 kDa band corresponding to Cofilin (pS3) was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Phospho-Cofilin (Ser3) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot analysis of Phospho-Cofilin (Ser3) using Phospho-Cofilin (Ser3) Rabbit Polyclonal Antibody (Product # 44-1072G) shows induced expression of Phospho-Cofilin (Ser3) in HeLa, A-431 and Jurkat cell lines treated with H2O2 and staurosporine respectively. {TM}

Product Details

44-1072G

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	-
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1:250	View 1 publication 1 publication

Product Specifications
Species Reactivity	Dog, Human, Mouse, Rat
Published species	Not Applicable
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human cofilin that contains serine 3. The sequence is conserved in mouse and rat. View immunogen
Conjugate	Unconjugated
Form	Liquid
Purification	Antigen affinity chromatography
Storage buffer	Dulbecco's PBS, pH 7.3, with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2533565

Target Information

Cofilin binds to F-actin and exhibits pH-sensitive F-actin depolymerizing activity. Regulates actin cytoskeleton dynamics. Important for normal progress through mitosis and normal cytokinesis. Plays a role in the regulation of cell morphology and cytoskeletal organization. Required for the up-regulation of atypical chemokine receptor ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation. Required for neural tube morphogenesis and neural crest cell migration.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Developmentally regulated GTP-binding protein 1 modulates ciliogenesis via an interaction with Dishevelled.

The Journal of cell biology

Lee M,Hwang YS,Yoon J,Sun J,Harned A,Nagashima K,Daar IO

44-1072G was used in Immunohistochemistry-immunofluorescence to show apical actin polymerization and stability in multiciliated cells is regulated by an interaction between developmentally regulated GTP-binding protein 1 and dishevelled.

Mon Aug 05 00:00:00 EDT 2019

Developmentally regulated GTP-binding protein 1 modulates ciliogenesis via an interaction with Dishevelled.

The Journal of cell biology

Lee M,Hwang YS,Yoon J,Sun J,Harned A,Nagashima K,Daar IO

Mon Aug 05 00:00:00 EDT 2019

Bioinformatics

Protein Aliases: 18 kDa phosphoprotein; CFL-1; COF1; cofilin 1 (non-muscle); Cofilin, non-muscle isoform; Cofilin-1; Cofilin1; epididymis secretory protein Li 15; p18

Gene Aliases: AA959946; CFL; CFL1; Cof; cofilin; HEL-S-15; n-cofilin

UniProt ID: (Human) P23528, (Rat) P45592, (Mouse) P18760

Entrez Gene ID: (Human) 1072, (Dog) 476022, (Rat) 29271, (Mouse) 12631

Molecular Function: non-motor actin binding protein