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Kit for detecting ER-alpha 36 expression level in tumour tissue

A technology of tumor tissue and expression level, which is applied in the field of medical devices and can solve problems such as non-standardized detection methods

Active Publication Date: 2019-04-09
BEIJING SHENOGEN BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, at present, there are no detection reagents or kits for the detection of ER-α36 in the world and domestically, and there is no corresponding standardized detection method

Method used

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  • Kit for detecting ER-alpha 36 expression level in tumour tissue
  • Kit for detecting ER-alpha 36 expression level in tumour tissue
  • Kit for detecting ER-alpha 36 expression level in tumour tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] See PCT / CN2014 / 073673, which is incorporated herein by reference.

[0049] The ER-α36 antibodies provided below include the SNGmB1 antibody.

[0050] The "SNGmB1 antibody" provided herein is a mouse-derived ER-α36 monoclonal antibody. It includes the light chain with the amino acid sequence of SEQ ID: 1 and the heavy chain with the amino acid sequence of SEQ ID NO: 2. The nucleotide sequences encoding the light chain and heavy chain of the SNGmB1 antibody are SEQ ID NO: 17 and SEQ ID NO: 19, respectively. The amino acid sequences of the light and heavy chains are shown below, with the CDR regions in italics and underlined, and the constant regions in italics shaded.

[0051] Amino acid sequence of SNGmB1 light chain (SEQ ID NO: 1):

[0052]

[0053]

[0054] Amino acid sequence of SNGmB1 heavy chain (SEQ ID NO: 2):

[0055]

[0056] Nucleotide sequence (SEQ ID NO: 9) of murine antibody SNGmB1 encoding the light chain:

[0057] 5‘-gaaacaactgtgacccagtctccagc...

Embodiment 2

[0065] The selection of reagent final concentration of the present invention

Embodiment 3

[0069] Obtain readable pathological slides by immunohistochemistry

[0070] fixed : First, soak the tumor tissue obtained by surgery or puncture in formalin fixative and fix it for 24 hours.

[0071] Washing and Dehydration : The fixed tissue is then gradually dehydrated with ethanol and absolute ethanol with concentration gradients of 50%, 70%, 85% and 95%.

[0072] Transparent : The dehydrated tissue is first soaked in the mixture of absolute ethanol:xylene=1:1 for 20 minutes, then transferred to pure xylene for 10 minutes, and then repeatedly immersed in another bottle of xylene for 10 minutes for proper treatment. transparent processing.

[0073] Waxing and embedding : After transparent treatment, immerse the tissue material in an equal mixture of molten paraffin and xylene for half an hour, then transfer to two melted paraffin liquids and immerse for one and a half hours respectively, and dip in the wax in an incubator at about 60°C conduct. The tissue materi...

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Abstract

The invention provides a kit for detecting the ER-alpha 36 expression level in tumour tissue. The kit comprises an ER-alpha 36 detection antibody reagent and a breast cancer tissue quality control table. The ER-alpha 36 expression level in breast cancer tissue of a patient is detected through the kit in the invention; therefore, prognosis after a breast cancer patient accepts tamoxifen treatment is judged in an assisted manner; and guidance significance is provided for clinic treatment.

Description

technical field [0001] The invention relates to a kit for detecting the expression level of ER-α36 in tumor tissue. The kit is used for detecting the expression level of ER-α36 in breast cancer tumor tissue and belongs to the field of medical equipment. Background technique [0002] Estrogen receptor ER-α36 is a new isomer of ER-α discovered and cloned in 2005 in human breast epithelial Caveolin-1 haploinsufficient cell line, with a molecular weight of 36kDa. ER-α36 differs from ER-α66 (the usual estrogen receptor) which is mainly present in the nucleus, this receptor is mainly distributed in the cytoplasm and / or cell membrane. The main difference between ER-α36 and ER-α66 is that it lacks two transcriptional activation sites, AF-1 and AF-2, but retains the DNA binding region and part of the dimer formation and ligand binding regions. ER-α36 functions similar to growth factor receptors, can interact with various estrogens, and participate in cell proliferation caused by est...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57415G01N33/57492G01N33/57496G01N33/68G01N2333/723
Inventor 孟坤陈凤白伟王伟娜郭兰英刘雅迪
Owner BEIJING SHENOGEN BIOMEDICAL
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