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  1. Link

    profilin-1-antibody-epr6304-ab124904.pdf

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Tags & Cell Markers Subcellular Markers Cytoskeleton Actin
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Profilin 1 antibody [EPR6304] (ab124904)

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  • SDS
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Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • OI-RD Scanning - Anti-Profilin 1 antibody [EPR6304] (ab124904)
  • Anti-Profilin 1 antibody [EPR6304] (ab124904)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6304] to Profilin 1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Profilin 1 antibody [EPR6304]
    See all Profilin 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR6304] to Profilin 1
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Profilin 1 aa 100-200 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat, JAR, HepG2 and HEK-293T cell lysates, mouse and rat brain lysate, human fetal brain lysate; IHC-P: Human colon and breast tissue; ICC/IF: HeLa, Jurkat cells; Flow Cyt (intra): HeLa cells;
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 1.24 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6304
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Actin
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Assembly
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Synapse marker
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Profilin 1 antibody [EPR6304] (ab203146)
    • Anti-Profilin 1 antibody [EPR6304] - BSA and Azide free (ab232020)
  • ChIP Related Products

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant Human Profilin 1 protein (ab87760)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124904 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (1)
1/10000 - 1/50000. Detects a band of approximately 12 kDa (predicted molecular weight: 15 kDa).
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.

ICC/IF
1/100.

For unpurified use at 1/250 - 1/500

Flow Cyt (Intra)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/10000 - 1/50000. Detects a band of approximately 12 kDa (predicted molecular weight: 15 kDa).
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.

ICC/IF
1/100.

For unpurified use at 1/250 - 1/500

Flow Cyt (Intra)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Binds to actin and affects the structure of the cytoskeleton. At high concentrations, profilin prevents the polymerization of actin, whereas it enhances it at low concentrations. By binding to PIP2, it inhibits the formation of IP3 and DG.
  • Sequence similarities

    Belongs to the profilin family.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Target information above from: UniProt accession P07737 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5216 Human
    • Entrez Gene: 18643 Mouse
    • Entrez Gene: 64303 Rat
    • Omim: 176610 Human
    • SwissProt: P07737 Human
    • SwissProt: P62962 Mouse
    • SwissProt: P62963 Rat
    • Unigene: 494691 Human
    • Unigene: 2647 Mouse
    • Unigene: 1152 Rat
    see all
  • Alternative names

    • Actin binding protein antibody
    • ALS18 antibody
    • Epididymis tissue protein Li 184a antibody
    • OTTHUMP00000125244 antibody
    • PFN 1 antibody
    • Pfn antibody
    • PFN1 antibody
    • PROF1_HUMAN antibody
    • Profilin I antibody
    • Profilin-1 antibody
    • Profilin1 antibody
    • ProfilinI antibody
    see all

Images

  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    All lanes : Anti-Profilin 1 antibody [EPR6304] (ab124904) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PFN1 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : Jurkat whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 15 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab124904 observed at 15 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab124904 was shown to specifically react with in wild-type HAP1 cells as signal was lost in PFN1 knockout cells. Wild-type and PFN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab124904 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling Profilin 1 with Purified ab124904 at 1:800 dilution (0.18 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Profilin 1 with Purified ab124904 at 1:100 dilution (1.4 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Anti-Profilin 1 antibody [EPR6304] (ab124904) at 1/10000 dilution (Purified) + Human fetal brain lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 15 kDa
    Observed band size: 12 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    All lanes : Anti-Profilin 1 antibody [EPR6304] (ab124904) at 1/10000 dilution (Purified)

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 15 kDa
    Observed band size: 12 kDa why is the actual band size different from the predicted?

  • Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Profilin 1 with Purified ab124904 at 1/200 dilution (1 µg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Western blot - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    All lanes : Anti-Profilin 1 antibody [EPR6304] (ab124904) at 1/10000 dilution (Unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : JAR cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 15 kDa
    Observed band size: 12 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Profilin 1 antibody [EPR6304] (ab124904)

    Unpurified ab124904, at 1/500 dilution, staining Profilin 1 in paraffin-embedded human colon tissue by Immunohistochemistry.

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Immunocytochemistry/ Immunofluorescence - Anti-Profilin 1 antibody [EPR6304] (ab124904)

    Unpurified ab124904, at 1/250 dilution, staining Profilin 1 in HeLa cells by Immunofluorescence.

  • Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Flow Cytometry (Intracellular) - Anti-Profilin 1 antibody [EPR6304] (ab124904)

    Overlay histogram showing HeLa cells stained with unpurifiedab124904 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab124904, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. 

     

     

  • OI-RD Scanning - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    OI-RD Scanning - Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-Profilin 1 antibody [EPR6304] (ab124904)
    Anti-Profilin 1 antibody [EPR6304] (ab124904)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (12)

Publishing research using ab124904? Please let us know so that we can cite the reference in this datasheet.

ab124904 has been referenced in 12 publications.

  • Zhu C  et al. Cancer-associated exportin-6 upregulation inhibits the transcriptionally repressive and anticancer effects of nuclear profilin-1. Cell Rep 34:108749 (2021). PubMed: 33596420
  • Gau D  et al. Disruption of profilin1 function suppresses developmental and pathological retinal neovascularization. J Biol Chem 295:9618-9629 (2020). PubMed: 32444495
  • Pan MH  et al. FMNL3 regulates FASCIN for actin-mediated spindle migration and cytokinesis in mouse oocytes†. Biol Reprod 102:1203-1212 (2020). PubMed: 32167535
  • Ren JY  et al. Protein Tyrosine Phosphatase Receptor S Acts as a Metastatic Suppressor in Malignant Peripheral Nerve Sheath Tumor via Profilin 1-Induced Epithelial-Mesenchymal Transition. Front Cell Dev Biol 8:582220 (2020). PubMed: 33163494
  • Zhang Y  et al. Loss of PKC mu function induces cytoskeletal defects in mouse oocyte meiosis. J Cell Physiol N/A:N/A (2019). PubMed: 30912144
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Profilin 1 antibody [EPR6304]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Heart)
Permeabilization
Yes - Tween-20
Specification
Embryonic Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 01 2016

Western blot abreview for Anti-Profilin 1 antibody [EPR6304]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 19 2016

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