Invitrogen

Profilin 1 Monoclonal Antibody (OTI1D5)

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

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2 篇参考文献

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Profilin 1 Antibody in Immunocytochemistry (ICC/IF)

Profilin 1 Antibody in Immunocytochemistry (ICC/IF)

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Profilin 1 Antibody (MA5-25113) in ICC/IF

Immunofluorescence of Profilin 1 (PFN1) was performed using HAP1 WT and PFN1 KO cells that were labeled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with glass bottom as a mosaic and incubated for 24 hrs. Cells were fixed in 4% PFA (in PBS) for 15 min at RT; cells were permeabilized with 0.1% Triton X-100 for 10 min at RT an... View More

Profilin 1 Antibody in Immunocytochemistry (ICC/IF)

Profilin 1 Antibody in Immunocytochemistry (ICC/IF)

Profilin 1 Antibody in Immunocytochemistry (ICC/IF)

Profilin 1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

Profilin 1 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot (WB)

Profilin 1 Antibody in Western Blot, Immunoprecipitation (WB, IP)

Profilin 1 Antibody

Profilin 1 Antibody

Profilin 1 Antibody

Product Image Gallery

Profilin 1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence of Profilin 1 (PFN1) was performed using HAP1 WT and PFN1 KO cells that were labeled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with glass bottom as a mosaic and incubated for 24 hrs. Cells were fixed in 4% PFA (in PBS) for 15 min at RT; cells were permeabilized with 0.1% Triton X-100 for 10 min at RT and blocked with PBS with 5% BSA, 5% goat serum, and 0.01% Triton X-100 for 30 min. Cells were stained with Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113) at a 1.0 µg/ml overnight at 4 degree celcius. Secondary antibody incubation was performed using 1 µg/mL of Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 555 (Product # A-21424) together with DAPI for 1 hr at RT. Imaging was performed with a 20X water immersion objective. Representative images where WT and KO cells are outlined with a green (WT) or magenta (KO) line, respectively, are shown. The top and bottom panels show antibody and DAPI stainings, respectively. Scale bar = 10 µm. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Profilin 1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Profilin 1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoskeleton, nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Profilin 1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of PFN1 in COS7 cells. Cells were transfected with a plasmid overexpressing PFN1 and probed with a PFN1 monoclonal antibody (Product # MA5-25113).

Profilin 1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded human pancreas tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a PFN1 monoclonal antibody (Product # MA5-25113) at a dilution of 1:50.

Profilin 1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded human kidney tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a PFN1 monoclonal antibody (Product # MA5-25113) at a dilution of 1:50.

Profilin 1 Antibody in Western Blot (WB) — Western blot analysis of PFN1 in human tissue (1: Testis; 2: Omentum; 3: Uterus; 4: Breast; 5: Brain; 6: Liver; 7: Ovary; 8: Thyroid gland; 9: colon;10: spleen) samples using 10 µg per lane. Samples were separated by SDS-PAGE and probed with PFN1 (Product # MA5-25113) monoclonal antibody at a dilution of 1:200.

Profilin 1 Antibody in Western Blot (WB) — Western blot analysis of PFN1 in HepG2, HeLa, HT29, A549, COS7, Jurkat, MDCK, PC12, MCF7 cells using 35 µg per lane. Samples were probed with PFN1 (Product # MA5-25113) monoclonal antibody.

Profilin 1 Antibody in Western Blot (WB) — Western blot was performed using Anti-Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113) and a 15kDa band corresponding to Profilin 1 was observed across cell lines. Whole cell extract (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), HEK-293 (Lane 3) and A-431 (Lane 4) were electrophoresed using Novex™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Profilin 1 Antibody in Western Blot (WB) — Western blot analysis of PFN1 in 3T3 cells, mouse brain, mouse liver, mouse spleen lysate using 10 µg per lane. Samples were probed with PFN1 (Product # MA5-25113) monoclonal antibody at a dilution of 1:200.

Profilin 1 Antibody in Western Blot (WB) — Western blot of Profilin 1 (PFN1) was performed by loading 50 µg of HAP1 WT (lane 1) and PFN1 KO (lane 2) cell lysates onto a NuPAGE™ 10%, Bis-Tris, 1.0 mm, Midi Protein Gel (Product # WG1201BOX)). Proteins on the blot were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. PFN1 was detected at approximately 15 kDa using Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113) at a dilution of 1:2000 in 5% BSA in TBST overnight at 4 degree celsius. The blot was probed with Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP (Product # 62-6520) diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr at room temperature. Chemiluminescent detection was performed using ECL Western Blotting Substrate. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Profilin 1 Antibody in Western Blot (WB) — Knockdown of Profilin 1 was achieved by transfecting HeLa cells with Profilin 1 specific siRNAs (Silencer® select Product # s10375, s10376). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Profilin 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Profilin 1.

Profilin 1 Antibody in Western Blot (WB) — Western blot analysis of PFN1 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with PFN1 (Product # MA5-25113) monoclonal antibody.

Profilin 1 Antibody in Western Blot (WB) — Figure 1. Profilin-1 antibody screening by Western blot. Lysates of HAP1 (WT and PFN1 KO) were prepared and 50 mug of protein were processed for Western blot with the indicated Profilin-1 antibodies. The Ponceau stained transfers of each blot are presented to show equal loading of WT and KO lysates and protein transfer efficiency from the polyacrylamide gels to the nitrocellulose membrane. Antibody dilutions were chosen according to the recommendations of the antibody supplier. An exception was given for antibody A9188** which was titrated to 1/30000, as the signal was too weak when following the supplier's recommendation. When the concentration was not indicated by the supplier, which was the case for AC-PFN1-4**, we tested the antibody at 1/1000. Antibody dilution used: 11680-1-AP at 1/6000, 67390-1-lg* at 1/30000, MAB7779* at 1/5000, NBP2-59778* at 1/1000, NBP2-67078* at 1/2000, GTX102072 at 1/3000, 3246** at 1/1000, A9188 at 1/30000, AC-PFN1-4** at 1/1000, ARP48269 at 1/2000, MA5-25113* at 1/2000, MA5-25120* at 1/2000, MA5-32683** at 1/2000, ab242369* at 1/2000, ab133529* at 1/10000, ab124904** at 1/30000. Predicted band size: 15 kDa. *Monoclonal antibody, **Recombinant antibody.

Profilin 1 Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 2. Profilin-1 antibody screening by immunoprecipitation. HAP1 lysates were prepared, and IP was performed using 2.0 mug of the indicated Profilin-1 antibodies pre-coupled to Dynabeads protein G or protein A or Flag-M2 magnetic beads. Samples were washed and processed for Western blot with the indicated Profilin-1 antibody. For Western blot, MA5-25120* was used at 1/5000. The Ponceau stained transfers of each blot are shown for similar reasons as in Figure 1 . SM=2% starting material; UB=2% unbound fraction; IP=Immunoprecipitate. *Monoclonal antibody, **Recombinant antibody.

Profilin 1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. Loss of signal in immunofluorescence was observed for target protein in Profilin 1 knockout (KO) cell line using Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113). Parental cells were labeled with CellTracker green (Product # C2925) and TARDBP KO with CellTracker Deep Red (Product # C34565). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation. {KO}

Profilin 1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with Profilin 1 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113). {KD}

Profilin 1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. Loss of signal was observed for target protein in Profilin 1 knockout (KO) cell line using Profilin 1 Monoclonal Antibody (OTI1D5) (Product # MA5-25113). Ponceau stained transfer shown below. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation. {KO}

产品信息

MA5-25113

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:1,000-1:2,000

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免疫组化（石蜡） (IHC (P))

1:50

-

免疫细胞化学 (ICC/IF)

1:100

-

流式细胞分析 (Flow)

1:100

-

免疫沉淀 (IP)

-

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产品规格

种属反应

Dog, Human, Non-human primate, Rat

已发表种属

Human

宿主/亚型

Mouse / IgG2a

分类

Monoclonal

类型

Antibody

克隆号

OTI1D5

抗原

Full length human recombinant protein of PFN1 produced in HEK293T cell

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

1 mg/mL

纯化类型

Affinity Chromatography

保存液

PBS, pH 7.3, with 1% BSA, 50% glycerol

内含物

0.02% sodium azide

保存条件

-20° C, Avoid Freeze/Thaw Cycles

运输条件

Ambient (domestic); Wet ice (international)

RRID

AB_2725111

靶标信息

Profilin is an ubiquitous small (12-15 kDa) phosphoinositides and poly-L-proline binding protein that plays a role in both signal transduction pathways and actin filament dynamics. There are two mammalian profilins with similar biochemical properties but different expression patterns. Profilin I appears to be highly expressed in most tissues except for skeletal muscle. Profilin II is predominantly expressed in brain and at lower levels in skeletal muscle, uterus and kidney. Profilin is a mainly cytosolic protein with higher concentrations in dynamic membrane areas like the leading edge and ruffling membranes.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

The identification of high-performing antibodies for Profilin-1 for use in Western blot, immunoprecipitation and immunofluorescence.

F1000Research

Ayoubi R,McDowell I,Fotouhi M,Southern K,McPherson PS,Laflamme C

MA5-25113 was used in Western Blot to select the most appropriate antibody for their specific needs.

Tue Aug 15 00:00:00 EDT 2023

The identification of high-performing antibodies for Profilin-1 for use in Western blot, immunoprecipitation and immunofluorescence.

F1000Research

Ayoubi R,McDowell I,Fotouhi M,Southern K,McPherson PS,Laflamme C

MA5-25113 was used in Western Blot to select the most appropriate antibody for their specific needs.

Tue Aug 15 00:00:00 EDT 2023

The identification of high-performing antibodies for Profilin-1 for use in Western blot, immunoprecipitation and immunofluorescence.

F1000Research

Ayoubi R,McDowell I,Fotouhi M,Southern K,McPherson PS,Laflamme C

MA5-25113 was used in Western Blot to select the most appropriate antibody for their specific needs.

Tue Aug 15 00:00:00 EDT 2023

生物信息学

蛋白别名: Epididymis tissue protein Li 184a; Profilin I; Profilin-1

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基因别名: ALS18; PFN1

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UniProt ID: (Human) P07737, (Rat) P62963

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Entrez Gene ID: (Dog) 607397, (Human) 5216, (Rat) 64303

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adenyl-nucleotide exchange factor activity actin binding actin monomer binding receptor binding protein binding phosphatidylinositol-4,5-bisphosphate binding Rho GTPase binding poly(A) RNA binding proline-rich region binding cadherin binding involved in cell-cell adhesion actin or actin-binding cytoskeletal protein cytoskeletal protein non-motor actin binding protein

参与通路

neural tube closure positive regulation of epithelial cell migration actin cytoskeleton organization negative regulation of actin filament polymerization positive regulation of actin filament polymerization negative regulation of actin filament bundle assembly positive regulation of actin filament bundle assembly positive regulation of ATPase activity positive regulation of transcription from RNA polymerase II promoter positive regulation of viral transcription protein stabilization positive regulation of DNA metabolic process positive regulation of stress fiber assembly negative regulation of stress fiber assembly Wnt signaling pathway, planar cell polarity pathway cellular response to growth factor stimulus cell-cell adhesion positive regulation of ruffle assembly regulation of transcription from RNA polymerase II promoter actin polymerization or depolymerization response to organic substance