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Cortactin Recombinant Rabbit Monoclonal Antibody [SC61-08]

Usd: 385

Specification

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Catalog# ET1610-87

Cortactin Recombinant Rabbit Monoclonal Antibody [SC61-08]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

Predicted reactivity

  • Rat

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Cortactin Recombinant Rabbit Monoclonal Antibody [SC61-08]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Cortactin aa 34-67 / 550.

Species Reactivity

Human, Mouse (Predicted: Rat)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 62 kDa

Positive Control

HAP1-parental cell lysate, Hela cell lysate, NIH/3T3 cell lysate, HeLa, human tonsil tissue.

Conjugation

unconjugated

Clone Number

SC61-08

RRID

AB_3069973

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • IF-Tissue

  • 1:100-1:500

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

Cortactin (also designated Ems-1) is a filamentous Actin (F-Actin) binding protein that has been shown to be a substrate for Src p60. Cortactin contains tandem 37 amino acid repeats at the amino-terminus and an SH3 domain at the carboxy-terminus. The tandem repeats appear to be necessary for F-Actin binding. Tyrosine phosphorylation of Cortactin by Src p60 results in diminished F-Actin binding to Cortactin and reduced F-Actin cross-linking activity. Cortactin has also been shown to be phosphorylated in response to FGF-1. Cortactin exhibits abundant expression in megakaryocytes and platelets, and it may play a role in the maturation of megakaryocytes.

Background References

1. Motonishi S et al. Sirtuin1 Maintains Actin Cytoskeleton by Deacetylation of Cortactin in Injured Podocytes. J Am Soc Nephrol 26:1939-59 (2015).

2. Kaur S et al. An unmet actin requirement explains the mitotic inhibition of clathrin-mediated endocytosis. Elife 3:e00829 (2014).

Post-translational Modification

Phosphorylated by PKN2 at both serine and threonine residues in a GTP-bound Rac1-dependent manner in hyaluronan-induced astrocytes and hence down-regulated CTTN ability to associates with filamentous actin (By similarity). Phosphorylated on tyrosine residues in response to CHRM1 activation (By similarity). Phosphorylated by PTK2/FAK1 in response to cell adhesion (By similarity). Phosphorylated by FER. Tyrosine phosphorylation in transformed cells may contribute to cellular growth regulation and transformation. Phosphorylated in response to FGR activation. Phosphorylation by SRC promotes MYLK binding.

Subcellular Location

Cytoplasm, Cell projection, Membrane.

UNIPROT

SwissProt: Q14247 Human

SwissProt: Q60598 Mouse

Unigene:107869 Rat

Synonyms

Amplaxin antibody

CTTN antibody

EMS 1 antibody

EMS1 antibody

FLJ34459 antibody

Mammary tumor and squamous cell carcinoma associated antibody

Oncogene EMS1 antibody

p80/85 src substrate antibody

Src substrate cortactin antibody

SRC8_HUMAN antibody

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Cortactin on different lysates with Rabbit anti-Cortactin antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Cortactin KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 62 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 40 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Cortactin on different lysates with Rabbit anti-Cortactin antibody (ET1610-87) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Cortactin KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 62 kDa
    Observed band size: 75 kDa

    Exposure time: 40 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-87) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Cortactin on different lysates with Rabbit anti-Cortactin antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/500 dilution.<br /><br />Lane 1: Hela cell lysate<br />Lane 2: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 62 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 30 seconds;<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cortactin on different lysates with Rabbit anti-Cortactin antibody (ET1610-87) at 1/500 dilution.

    Lane 1: Hela cell lysate
    Lane 2: NIH/3T3 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 62 kDa
    Observed band size: 75 kDa

    Exposure time: 30 seconds;

    8% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-87) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Cortactin with Rabbit anti-Cortactin antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cortactin antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Cortactin with Rabbit anti-Cortactin antibody (ET1610-87) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cortactin antibody (ET1610-87) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Cortactin antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Cortactin antibody (ET1610-87) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-87) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling Cortactin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1610-87" style="font-weight: bold;text-decoration: underline;">ET1610-87</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Cortactin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-87, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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