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Application of CKAP4 reagent in detection specimen and bladder cancer detection kit

A bladder cancer and kit technology, applied in the field of tumor molecular biology, can solve problems such as poor signal, insufficient detection sensitivity, and inability to achieve statistical differences, and achieve the effect of avoiding missed detection

Pending Publication Date: 2022-05-31
HUNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, in the study of exosome biomarkers, TACSTD2, AAT, Filamin-A and H2B1K were found to be high in tumor-derived exosome samples, but detected by specific antibody blotting techniques, the signals were found to be poor, The statistical difference between the patients in each group and the control samples is still unable to be achieved, and the detection sensitivity is insufficient, making it difficult to translate into clinical application results

Method used

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  • Application of CKAP4 reagent in detection specimen and bladder cancer detection kit
  • Application of CKAP4 reagent in detection specimen and bladder cancer detection kit
  • Application of CKAP4 reagent in detection specimen and bladder cancer detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Tissue microarrays containing normal, malignant and metastases were purchased from Avilabio (Xi'an, China, DC-Bla11021). Will

Embodiment 2

[0097] Example 2: Using gradient centrifugation to collect exosomes in cell samples.

[0107] Loading: the prepared sample is added to the SDS-PAGE gel, and the electrophoresis program is 70V 35min, 120V 90min. Transfer film

[0109] (10) The antibody was recovered and washed three times with TBST for 10 min each time. Incubate mouse and rabbit antibodies at room temperature for 1 h. Recovered Antibody, TBST

[0112] (1) 5× electrophoresis buffer: 288g, Gly; 60.6g, Tris; 20g, SDS, and finally add deionized water to dilute to 4L. Make

[0113] (2) Transfer liquid: 14.4g, glycine; 3.03g, Tris; 200m, methanol, and finally add deionized water and make up to 1L.

[0117] 10% separating gel and 5% stacking gel. 10% SDS‑PAGE Protein Separation Gel (5mL): In a 50mL centrifuge tube, press

[0119] Transmission electron microscopy (TEM) was used to observe the exosomes of 5637 and SV-HUC-1 cells. will first be able to specifically identify

[0120] The results are shown in Figure 2.

Embodiment 3

[0124] MPSAKQRGSKGGHGAASPSEKGAHPSGGADDVAKKPPPAPQQPPPPPAPHPQQHPQQHPQNQAHGKGGH

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Abstract

The invention discloses application of a reagent for detecting CKAP4 in a sample and a bladder cancer detection kit. The CKAP4 is not only highly expressed in bladder cancer cell lines and exosomes secreted by the bladder cancer cell lines, but more importantly, the CKAP4 is remarkably and highly expressed in urine exosomes of bladder cancer patients clinically, but the expression quantity in urinary system cancers such as prostatic cancer and renal cell carcinoma, urinary system inflammation and urine exosomes of healthy donors is very low, and the CKAP4 has remarkable specificity. Therefore, the detection means based on the urine exosome CKAP4 is a non-invasive liquid biopsy method which is low in cost, simple and convenient, can meet the requirement for multiple times of detection, and is particularly suitable for diagnosis and relapse monitoring of bladder cancer patients.

Description

Application of CKAP4 reagent in detection specimen and bladder cancer detection kit technical field The invention belongs to tumor molecular biology technical field, relate to a kind of application of CKAP4 reagent in detection specimen and Bladder Cancer Detection Kit. Background technique [0002] Bladder cancer is a malignant urological disease that occurs in the bladder epithelium. According to 2020 data, the global Bladder cancer is in the top 10 for both incidence and mortality, with 573,000 new cases and 213,000 deaths annually. Of these, approximately 75% are non-muscle-invasive bladder cancers and 25% are muscle-invasive bladder cancers. However, whether or not Muscle invasion, an important feature of bladder cancer is its high recurrence, and metastasis of bladder cancer can cause rapid disease progression. development, and the patient died in a short period of time. Therefore, it is necessary to establish a fast, simple, and inexpensive method that can m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57407G01N33/57488G01N33/57484G01N33/6854G01N2333/47
Inventor 叶茂谭蔚泓孙星李霞辉
Owner HUNAN UNIV
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