ETV4 ELISA Kit

Name: ETV4 ELISA Kit
SKU: DEIA-XYA631
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

nuclear, cell lysates

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The ETV4 ELISA Kit is used for Detection and Qualitative Analysis of Activated Transcription Factors in Nuclear Lysates, Cytoplasmic Lysates and Tissue Homogenates

Contents of Kit

1. 96-Well dsDNA Oligonucleotide Coated Microplate, 12×8 wells
2. 100× Primary Antibody, 100 μl
3. HRP-Conjugated Anti-Rabbit lgG Antibody, 12 ml
4. Nuclear Lysate Positive Control, Lyophilized
5. Wild-Type Consensus dsDNA Oligonuceotide, 10 μl
6. Mutant Consensus dsDNA Oligonucleotide, 10 μl
7. 10× Wash Buffer, 50 ml
8. 2× Binding Buffer, 12 ml
9. Primary Antibody Diluent, 12 ml
10. Nuclear Wash Buffer, 12 ml
11. Cytoplasmic Extraction Buffer, 6 ml
12. Nuclear Extraction Buffer, 6 ml
13. Ready-to-Use Substrate, 12 ml
14. Stop Solution, 12 ml
15. Adhesive Plate Sealers
16. Technical Manual

Storage

Note: After receiving the the kit for the first time, please store the kit components at the temperature indicated in the table below.
Note: After using the kit the for the first time the storage times are indicated below. Reconstituted Nuclear Lysate Positive Control must be aliquoted and stored at -80°C.
1. 96-Well dsDNA Oligonucleotide Coated Microplate, 1 month, 4°C
2. 100× Primary Antibody, 1 month, 4°C
3. HRP-Conjugated Anti-Rabbit lgG Antibody, 1 month, 4°C
4. Nuclear Lysate Positive Control, Lyophilized: 1 year 4°C, Reconstituted: 1 month -80°C.
5. Wild-Type Consensus dsDNA Oligonuceotide, 1 month, -20°C
6. Mutant Consensus dsDNA Oligonucleotide, 1 month, -20°C
7. 10× Wash Buffer, 1 month, 4°C
8. 2× Binding Buffer, 1 month, -20°C
9. Primary Antibody Diluent, 1 month, 4°C
10. Nuclear Wash Buffer, 1 month, -20°C
11. Cytoplasmic Extraction Buffer, 1 month, -20°C
12. Nuclear Extraction Buffer, 1 month, -20°C
13. Ready-to-Use Substrate, 1 month, 4°C
14. Stop Solution, 1 month, 4°C

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

CIC break-apart fluorescence in-situ hybridization misses a subset of CIC-DUX4 sarcomas: a clinicopathological and molecular study

HISTOPATHOLOGY

Authors: Yoshida, Akihiko; Arai, Yasuhito; Kobayashi, Eisuke; Yonemori, Kan; Ogura, Koichi; Hama, Natsuko; Mukai, Wakako; Motoi, Toru; Kawai, Akira; Shibata, Tatsuhiro; Hiraoka, Nobuyoshi

Abstract

AimsApproximately 60-70% of high-grade round-cell sarcomas that lack the Ewing sarcoma breakpoint region 1 (EWSR1) rearrangement harbour a rearrangement of the CIC gene, most commonly CIC-DUX4. Recent studies have established that CIC-rearranged sarcomas constitute a distinct group characterized by recognizable histology and immunoprofiles, such as positivity for ETV4 and WT1 and negativity for NKX2.2. Although these sarcomas are diagnosed increasingly in practice by fluorescence in-situ hybridization (FISH) with CIC break-apart probes, the optimal modality to diagnose these sarcomas has not been determined. In this study, we describe four round-cell sarcomas that showed false-negative results by CIC break-apart FISH assays. Methods and resultsThese sarcomas showed characteristic histology of CIC-rearranged sarcomas, and all were immunohistochemically positive for ETV4 and WT1 and negative for NKX2.2. Although FISH showed non-atypical negative signals for CIC rearrangement, high-throughput RNA sequencing identified CIC-DUX4 and its fusion breakpoint in all cases. Their clinical and histological findings, as well as fusion points determined by RNA sequencing, did not differ significantly from those of nine FISH-positive CIC-DUX4 sarcoma cases. We estimated that the FISH false-negative rate for CIC-rearranged sarcomas was 14%. Although neither histology nor immunoprofiles (e.g. ETV4 and WT1) are entirely sensitive or specific for CIC-rearranged sarcomas, the observation that these four cases were identified successfully by such phenotypes suggested their practical utility. ConclusionsCIC break-apart FISH assays missed a significant minority of CIC-DUX4 sarcomas, and full awareness of typical morphology and judicious immunohistochemical work-ups, including analyses of ETV4 and WT1, should complement diagnostic assessment.

Capicua restricts cancer stem cell-like properties in breast cancer cells

ONCOGENE

Authors: Yoe, Jeehyun; Kim, Donghyo; Kim, Sanguk; Lee, Yoontae

Abstract

Cancer stem cells (CSCs) play a central role in cancer initiation, progression, therapeutic resistance, and recurrence in patients. Here we present Capicua (CIC), a developmental transcriptional repressor, as a suppressor of CSC properties in breast cancer cells. CIC deficiency critically enhances CSC self-renewal and multiple CSC subpopulations of breast cancer cells without altering their growth rate or invasiveness. Loss of CIC relieves repression of ETV4 and ETV5 expression, consequently promoting self-renewal capability, EpCAM(+)/CD44(+)/CD24(low/-) expression, and ALDH activity. In xenograft models, CIC deficiency significantly increases CSC frequency and drives tumor initiation through derepression of ETV4. Consistent with the experimental data, the CD44(high)/CD24(low) CSC-like feature is inversely correlated with CIC levels in breast cancer patients. We also identify SOX2 as a downstream target gene of CIC that partly promotes CSC properties. Taken together, our study demonstrates that CIC suppresses breast cancer formation via restricting cancer stemness and proposes CIC as a potential regulator of stem cell maintenance.