ECT2 Polyclonal antibody

ECT2 Polyclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Cat no : 26557-1-AP

Synonyms

Protein ECT2, Epithelial cell-transforming sequence 2 oncogene

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Validation Data Gallery

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  WB analysis using 26557-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 26557-1-AP (ECT2 antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  WB analysis using 26557-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 26557-1-AP (ECT2 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of K-562 using 26557-1-AP

  IP result of anti-ECT2 (IP:26557-1-AP, 4ug; Detection:26557-1-AP 1:6000) with K-562 cells lysate 2400 ug.

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  IF Staining of A549 using 26557-1-AP

  Immunofluorescent analysis of (4% PFA) fixed A549 cells using ECT2 antibody (26557-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

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  IF Staining of NIH/3T3 using 26557-1-AP

  Immunofluorescent analysis of (4% PFA) fixed NIH/3T3 cells using ECT2 antibody (26557-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of A549 using 26557-1-AP

  1x10^6 A549 cells were intracellularly stained with 0.25 ug ECT2 Polyclonal antibody (26557-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: A549 cells, HEK-293T cells, K-562 cells
• Positive IP detected in: K-562 cells
• Positive IF/ICC detected in: A549 cells, NIH/3T3 cells
• Positive FC (Intra) detected in: A549 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:3000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 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.

Product Information

26557-1-AP targets ECT2 in WB, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: ECT2 fusion protein Ag24836
• Full Name: epithelial cell transforming sequence 2 oncogene
• Observed Molecular Weight: 100 kDa
• GenBank Accession Number: BC112086
• Gene Symbol: ECT2
• Gene ID (NCBI): 1894
• RRID: AB_3669547
• 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

Epithelial cell transforming sequence 2 (Ect2) is a guanine nucleotide exchange factor (GEF) for the Rho family GTPases, Rac1, RhoA and Cdc42. Ect2 catalyzes the exchange of GDP for GTP, thereby activating the Rho GTPases toward their downstream effectors  (PMID: 35069543). The cellular localization of Ect2 is cell cycle dependent. Ect2 is localized predominantly in the nucleus during interphase due to two functional NLS sequences found within its central hinge region.1 At the onset of mitosis, Ect2 is dispersed throughout the cytoplasm where it associates with the mitotic spindle at metaphase, the cleavage furrow during telophase, and the mid-body during cytokinesis (PMID: 35069543). Besides normal cellular activity, ECT2 also participates in malignant transformation, tumor initiation, and metastasis (PMID: 36572949).