JPH0746103B2 - Method for directly measuring thiol group in serum and reagent used therefor - Google Patents
Method for directly measuring thiol group in serum and reagent used thereforInfo
- Publication number
- JPH0746103B2 JPH0746103B2 JP31747987A JP31747987A JPH0746103B2 JP H0746103 B2 JPH0746103 B2 JP H0746103B2 JP 31747987 A JP31747987 A JP 31747987A JP 31747987 A JP31747987 A JP 31747987A JP H0746103 B2 JPH0746103 B2 JP H0746103B2
- Authority
- JP
- Japan
- Prior art keywords
- serum
- thiol group
- directly measuring
- reagent used
- present
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 210000002966 serum Anatomy 0.000 title claims description 39
- 125000003396 thiol group Chemical group [H]S* 0.000 title claims description 31
- 238000000034 method Methods 0.000 title claims description 22
- 239000003153 chemical reaction reagent Substances 0.000 title claims description 20
- 238000005259 measurement Methods 0.000 claims description 9
- 108090000623 proteins and genes Proteins 0.000 claims description 9
- 102000004169 proteins and genes Human genes 0.000 claims description 9
- 229960000789 guanidine hydrochloride Drugs 0.000 claims description 8
- YLZSIUVOIFJGQZ-UHFFFAOYSA-N bis[4-(dimethylamino)phenyl]methanol Chemical compound C1=CC(N(C)C)=CC=C1C(O)C1=CC=C(N(C)C)C=C1 YLZSIUVOIFJGQZ-UHFFFAOYSA-N 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical group [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims description 5
- 239000003398 denaturant Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 description 9
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 7
- 108010088751 Albumins Proteins 0.000 description 7
- 102000009027 Albumins Human genes 0.000 description 7
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 238000004440 column chromatography Methods 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 238000011088 calibration curve Methods 0.000 description 4
- 108010004546 mercaptoalbumin Proteins 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 230000002452 interceptive effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 2
- HFZDHVOXZVBBNW-UHFFFAOYSA-N C1(=CC=C(C=C1)N(C1=CC=C(C=C1)N=NC1=CC=CC=C1)C1=CC=C(C=C1)C1=CC=CC=C1)C1=CC=CC=C1 Chemical compound C1(=CC=C(C=C1)N(C1=CC=C(C=C1)N=NC1=CC=CC=C1)C1=CC=C(C=C1)C1=CC=CC=C1)C1=CC=CC=C1 HFZDHVOXZVBBNW-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 238000000691 measurement method Methods 0.000 description 2
- PVOAHINGSUIXLS-UHFFFAOYSA-N 1-Methylpiperazine Chemical compound CN1CCNCC1 PVOAHINGSUIXLS-UHFFFAOYSA-N 0.000 description 1
- SLAMLWHELXOEJZ-UHFFFAOYSA-N 2-nitrobenzoic acid Chemical compound OC(=O)C1=CC=CC=C1[N+]([O-])=O SLAMLWHELXOEJZ-UHFFFAOYSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- PKKSAVQQCBTBNZ-UHFFFAOYSA-N amino(diphenyl)methanol Chemical compound C=1C=CC=CC=1C(O)(N)C1=CC=CC=C1 PKKSAVQQCBTBNZ-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000020832 chronic kidney disease Diseases 0.000 description 1
- 208000022831 chronic renal failure syndrome Diseases 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
- 229940043264 dodecyl sulfate Drugs 0.000 description 1
- 238000003370 dye binding method Methods 0.000 description 1
- 238000007336 electrophilic substitution reaction Methods 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 238000007344 nucleophilic reaction Methods 0.000 description 1
- 208000030212 nutrition disease Diseases 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000034005 thiol-disulfide exchange Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Description
【発明の詳細な説明】 ＜産業上の利用分野＞ この発明は、血清中のチオール基、特に血清中のメルカ
プトアルブミンを主体とするチオール基をクロマトグラ
フィー等でメルカプトアルブミン成分を分画することな
く直接測定（定量）する方法、及びこれに用いる試薬に
関するものである。DETAILED DESCRIPTION OF THE INVENTION <Industrial field of application> The present invention is directed to a thiol group in serum, particularly a thiol group mainly composed of mercaptoalbumin in serum without fractionating a mercaptoalbumin component by chromatography or the like. The present invention relates to a method for direct measurement (quantification) and a reagent used therefor.
＜従来の技術＞ 血清中のアルブミン量と疾患との関係は、特に栄養障害
時の有効な指標とされるので、アルブミン量はこれと親
和性を有する色素を使用して、色素結合法という簡便法
によって日常的に測定されている。<Prior Art> Since the relationship between the amount of albumin in serum and disease is an effective index especially in the case of nutritional disorders, the amount of albumin is determined by a dye binding method using a dye having an affinity with this. It is routinely measured by law.
他方、アルブミン分子に存在するチオール基について
は、慢性腎不全、ネフローゼ症候群、各種肝疾患等によ
り、これがシステインやグルタチオン等と結合し、その
結果Ｓ−Ｓ（混合ジスルフィド）結合したアルブミンが
増大することが報告されている。従ってチオール基を持
つメルカプトアルブミン（以下HMAという）と疾患によ
り増えるノンメルカプトアルブミン（以下HNAという）
の含有率を測定することは、臨床検査の分野において有
用なことである。On the other hand, regarding the thiol group present in the albumin molecule, due to chronic renal failure, nephrotic syndrome, various liver diseases, etc., this binds with cysteine, glutathione, etc., and as a result, SS (mixed disulfide) bound albumin increases. Has been reported. Therefore, mercaptoalbumin with a thiol group (hereinafter referred to as HMA) and non-mercaptoalbumin (hereinafter referred to as HNA) that increases due to disease
It is useful in the field of clinical examination to measure the content rate of.
さて、血清中には種々の干渉物質が存在するので、従来
血清中のチオール基（特に血清中のHMAを主体とするチ
オール基）を単に試薬を添加するだけで直接測定するこ
とはできななった。Since there are various interfering substances in serum, it has not been possible to directly measure thiol groups in serum (especially thiol groups mainly composed of HMA in serum) in conventional serum simply by adding reagents. It was
すなわち、血清中のチオール基の測定は、その中に含ま
れているアルブミンを予じめカラムクロマトグラフィー
等により分画して共存する干渉物質を除去した後、これ
に5,5′−ジチオビスコニトロベンゾイックアシッド（D
TNB）を添加してアルブミンのチオール基により呈色さ
せ、その吸光度を測定することにより行なわれていたの
である。That is, the measurement of the thiol group in serum was carried out by preliminarily fractionating albumin contained therein by column chromatography or the like to remove coexisting interfering substances, and then adding 5,5'-dithiobisco Nitrobenzoic acid (D
It was carried out by adding TNB) to cause coloration by the thiol group of albumin and measuring the absorbance.
＜発明が解決しようとする問題点＞ しかしながら、前記したカラムクロマトグラフィー等に
より分画する操作は煩雑で時間がかかり過ぎるので、血
清中のチオール基の直接測定方法の開発が望まれてい
た。<Problems to be Solved by the Invention> However, since the operation of fractionating by the above-mentioned column chromatography and the like is complicated and takes too much time, development of a method for directly measuring a thiol group in serum has been desired.
発明者等は、前記DTNBを使用して、血清中のチオール基
を直接測定する方法について、カラムクロマトグラフィ
ー法と比較検討したところ、DTNBの呈色を阻害するの
は、血清中の低分子物質（セファデックスＧ−25脱塩用
カラムの低分子分画）である事実を突きとめたが、いず
れにしても従来血清中のチオール基の測定試薬として使
用されていたDTNBは、チオール基の直接測定試薬として
は不適当であることが判明した（第１図参照）。The inventors of the present invention, using the DTNB, a method for directly measuring the thiol group in serum, as a result of comparative examination with a column chromatography method, inhibits the coloration of DTNB is a low molecular weight substance in serum. We have identified the fact that it is a (low molecular weight fraction of Sephadex G-25 desalting column), but in any case, DTNB, which was conventionally used as a reagent for measuring thiol groups in serum, directly It was found to be unsuitable as a measuring reagent (see Fig. 1).
そこで発明者等は、DTNBとは異なった機構（反応）によ
り呈色する試薬を鋭意検討した結果、血清中のチオール
基の直接測定試薬（呈色試薬）としては、4,4′−ビス
ジメチルアミノベンズヒドロール（以下BDABという）が
最適であること、また血清中の総チオール基量はHMAで
大部分を占められているという事実を知り本発明を完成
した。Therefore, the present inventors have diligently studied a reagent that develops a color by a mechanism (reaction) different from DTNB, and as a reagent for directly measuring a thiol group in serum (color reagent), 4,4′-bisdimethyl The present invention has been completed by knowing that aminobenzhydrol (hereinafter referred to as BDAB) is optimal and that the total amount of thiol groups in serum is dominated by HMA.
＜問題を解決するための手段＞ すなわちこの発明は、 血清に、4,4′−ビスジメチルアミノベンズヒドロー
ル、及び蛋白変性剤を添加して生ずる呈色の程度を比較
測定することを特徴とする血清中のチオール基の直接測
定方法、及び 4,4′−ビスジメチルアミノベンズヒドロール（BDA
B）及び蛋白変性剤を含有することを特徴とする血清中
のチオール基の直接測定方法に用いる試薬、 を提供し、血清中から予じめアルブミンの分画しなくて
もチオール基を直接測定することができるようにするこ
とを目的として開発したものである。<Means for Solving Problems> That is, the present invention is characterized in that the degree of coloration produced by adding 4,4′-bisdimethylaminobenzhydrol and a protein denaturant to serum is comparatively measured. Method for direct measurement of thiol groups in serum, and 4,4'-bisdimethylaminobenzhydrol (BDA
B) and a reagent used in a method for directly measuring a thiol group in serum characterized by containing a protein denaturing agent, and directly measuring a thiol group without preliminarily fractionating albumin from serum It was developed with the purpose of making it possible.
本発明においては、血清に4,4′−ビスジメチルアミノ
ベンズヒドロールと蛋白変性剤を添加するものである
が、他の物質を添加しないという趣旨ではない。分析
上、通常必要とする他の物質を添加することはもちろん
である。In the present invention, 4,4'-bisdimethylaminobenzhydrol and a protein denaturant are added to serum, but it does not mean that other substances are not added. Of course, other substances usually required for analysis are added.
本発明に使用するBDABは、従来のDTNBとは次に記載する
ようにチオール基に対する特性が異なっている。BDAB used in the present invention is different from conventional DTNB in properties with respect to thiol groups as described below.
すなわち、従来のDTNB法によるチオール−ジスルフィド
交換反応が親電子置換反応であるのに対し、本発明にお
けるBDAB法は、以下に示すようにカチオンに対するチオ
ール基の求核反応の結果生ずる呈色（吸光度変化）をモ
ニターする点にある。That is, while the thiol-disulfide exchange reaction by the conventional DTNB method is an electrophilic substitution reaction, the BDAB method in the present invention, as shown below, the coloration (absorbance resulting from the nucleophilic reaction of a thiol group with respect to a cation). Change).
従って、血清等の体液中に共存する低分子干渉物質の影
響を受けないので、カラムクロマトグラフィー等による
前処理を必要とせず、直接測定が可能となるのである。 Therefore, since it is not affected by a low-molecular interfering substance that coexists in a body fluid such as serum, direct measurement can be performed without requiring pretreatment such as column chromatography.
本発明に係る血清中のチオール基の直接測定方法及びそ
れに用いる試薬には、蛋白変性剤を必要とする。蛋白変
性剤としては、塩酸グアニジンが最も適している。その
他、ラウリル硫酸、尿素、エタノール、アセトン等も使
用できる。The method for directly measuring a thiol group in serum according to the present invention and the reagent used therefor require a protein denaturing agent. Guanidine hydrochloride is the most suitable protein denaturant. In addition, lauryl sulfate, urea, ethanol, acetone and the like can be used.
塩酸グアニジンが測定系に存在すると、１×105という
高い分子吸光係数を得られるため、検体は極めて少量で
よい。When guanidine hydrochloride is present in the measurement system, a high molecular extinction coefficient of 1 × 10 5 can be obtained, and therefore a very small amount of sample is required.
また、分光光度計で測定する際の吸収極大が612nmと長
波長側にあるので共存物質の影響を全く受けないことが
有利である。Further, since the absorption maximum at the time of measurement with a spectrophotometer is on the long wavelength side of 612 nm, it is advantageous that it is not affected by coexisting substances at all.
なお、本願に係る血清中のチオール基の直接測定方法及
びそれに用いる試薬は、測定対象が血清ばかりではな
く、他の体液にも応用可能である。The method for directly measuring a thiol group in serum and the reagent used therefor according to the present application can be applied not only to serum but also to other body fluids.
＜実施例１＞ 試薬:6M−塩酸グアニジン、14μＭ−DBABを拭む50mM−
酢酸緩衝液（ph5.0） 操作：検体50μに、前記試薬2mlを添加して十分撹拌
後、室温で20分間放置し、波長612nmで吸光度を測定す
る。同時にHMAの希釈列を検体と同様に操作して、その
濃度と得られた吸光度から検量線を作成し、血清中の総
チオール基物質量を求めた。得られた検量線を第２図
（Ａ）に示す。<Example 1> Reagents: 6M-guanidine hydrochloride, 14 μM-50 mM wiped with DBAB-
Acetate buffer (ph5.0) procedure: Add 2 ml of the above reagent to 50 μm of a sample, stir well, leave at room temperature for 20 minutes, and measure the absorbance at a wavelength of 612 nm. At the same time, the HMA dilution series was operated in the same manner as the sample, and a calibration curve was prepared from the concentration and the obtained absorbance to determine the total amount of thiol group substances in serum. The obtained calibration curve is shown in FIG.
また、6M−塩酸グアニジンに代えて0.17M−ラウリル硫
酸ナトリウムを使用したときの検量線を第２図（Ｂ）に
示す。A calibration curve obtained when 0.17M-sodium lauryl sulfate was used instead of 6M-guanidine hydrochloride is shown in FIG. 2 (B).
次に、健常者及び各疾患者（リウマチ、痛風、糖尿病、
癌）の血清を用いて、本発明に係る血清中のチオール基
の直接測定方法を実施した。また、比較としてHPLC（高
速液体カラムクロマトグラフィー，旭化成（株）製 ア
サヒパックES−502のIEC,移動相として0.4M 硫酸ナト
リウム,0.05M Ｎ−メチルピペラジン,1％ シアン化メ
チル（pH4.8））で血清を分離し、HMA及びHNAを測定し
た。またHMA/（HMA＋HNA）を求めた。その結果を第３図
及び第４図に示す。Next, healthy people and people with various diseases (rheumatism, gout, diabetes,
The direct measurement method of thiol group in serum according to the present invention was carried out using serum of cancer). For comparison, HPLC (high-performance liquid column chromatography, Asahi Kasei Corp. Asahi Pack ES-502 IEC, mobile phase 0.4 M sodium sulfate, 0.05 M N-methylpiperazine, 1% methyl cyanide (pH 4.8)) ), Serum was separated and HMA and HNA were measured. Further, HMA / (HMA + HNA) was calculated. The results are shown in FIGS. 3 and 4.
第３図及び第４図によれば、従来のHPLC法による疾患の
検定法よりも、本願発明の方法による疾患の検定方法の
方が有意性が高いこと、すなわち健常者及び各疾患者の
差が明らかである。According to FIGS. 3 and 4, the significance of the disease assay method according to the method of the present invention is higher than that of the conventional HPLC disease assay, that is, the difference between healthy subjects and individual diseased subjects. Is clear.
本発明に係る血清中のチオール基の直接測定方法及びそ
れに用いる試薬について、試薬の濃度、測定条件等を補
足すると次の通りである。Regarding the method for directly measuring a thiol group in serum according to the present invention and the reagent used therefor, the concentration of the reagent, measurement conditions and the like will be supplemented as follows.
（１）前記した試薬は塩酸グアニジンにおいては２〜６
モル、DBABにおいては、４〜20μＭの範囲で使用可能で
ある。(1) The above reagents are 2 to 6 in guanidine hydrochloride.
It can be used in the range of 4 to 20 μM in molar and DBAB.
（２）DBABの緩衝液の至適pHは5.0である。(2) The optimum pH of the DBAB buffer solution is 5.0.
（３）試薬を添加して10分後には、吸光度がほぼ安定す
る。(3) The absorbance is almost stable 10 minutes after the addition of the reagent.
＜発明の効果＞ 以上のように、この発明に係る血清中のチオール基の直
接測定方法及びそれに用いる試薬によれば、血清中のHM
Aを主とするチオール基含有量を、HPLC等により分画す
ることなく直接測定することがな可能となるので、日常
の検査に迅速、簡便かつ高精度の測定法として十分利用
できる。<Effect of the Invention> As described above, according to the method for directly measuring a thiol group in serum and the reagent used therefor according to the present invention, HM in serum is
Since the thiol group content mainly containing A can be directly measured without fractionation by HPLC or the like, it can be sufficiently used as a quick, simple and highly accurate measurement method for daily inspection.
また、これを用いた疾患の検定は、従来のHPLC法による
HMAとHNAの含有比率による検定よりも有意性が高いとい
う効果を有する。In addition, the disease assay using this is based on the conventional HPLC method.
It has the effect of being more significant than the test based on the content ratio of HMA and HNA.
第１図は、DTNBを使用する、血清中のHMAの直接測定結
果と、血清中の低分子物質（セファデックスＧ−25脱塩
用カラムの低分子分画）を除去したときの測定結果を示
す。 第２図（Ａ）、（Ｂ）は、本願発明によるHMAの検量線
（各々順に、6M−塩酸グアニジン、及び0.17M−ラウリ
ル硫酸ナトリウムを使用）示す。 第３図は、HPLC法によるHMA/（HMA＋HNA）を示す。 第４図は、本願発明の血清中のチオール基の直接測定方
法による総SH基濃度を示す。Fig. 1 shows the results of direct measurement of HMA in serum using DTNB and the results of removal of low molecular substances in serum (low molecular fraction of Sephadex G-25 desalting column). Show. FIGS. 2 (A) and 2 (B) show the calibration curve of HMA according to the present invention (6M-guanidine hydrochloride and 0.17M-sodium lauryl sulfate were used, respectively). FIG. 3 shows HMA / (HMA + HNA) by HPLC method. FIG. 4 shows the total SH group concentration by the method for directly measuring thiol groups in serum of the present invention.
Claims (4)
ズヒドロール、及び蛋白変性剤を添加して生ずる呈色の
程度を比較測定することを特徴とする血清中のチオール
基の直接測定方法。1. A direct measurement of thiol groups in serum, which comprises comparatively measuring the degree of coloration produced by adding 4,4'-bisdimethylaminobenzhydrol and a protein denaturant to serum. Method.
求の範囲第１項記載の血清中のチオール基の直接測定方
法。2. The method for directly measuring a thiol group in serum according to claim 1, wherein the protein denaturing agent is guanidine hydrochloride.
ール及び蛋白変性剤を含有することを特徴とする血清中
のチオール基の直接測定方法に用いる試薬。3. A reagent used in a method for directly measuring a thiol group in serum, which comprises 4,4'-bisdimethylaminobenzhydrol and a protein denaturing agent.
求の範囲第３項記載の血清中のチオール基の直接測定方
法に用いる試薬。4. The reagent used in the method for directly measuring a thiol group in serum according to claim 3, wherein the protein denaturing agent is guanidine hydrochloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP31747987A JPH0746103B2 (en) | 1987-12-17 | 1987-12-17 | Method for directly measuring thiol group in serum and reagent used therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP31747987A JPH0746103B2 (en) | 1987-12-17 | 1987-12-17 | Method for directly measuring thiol group in serum and reagent used therefor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01161151A JPH01161151A (en) | 1989-06-23 |
| JPH0746103B2 true JPH0746103B2 (en) | 1995-05-17 |
Family
ID=18088685
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP31747987A Expired - Fee Related JPH0746103B2 (en) | 1987-12-17 | 1987-12-17 | Method for directly measuring thiol group in serum and reagent used therefor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0746103B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0788595B1 (en) * | 1994-10-27 | 2004-06-23 | Campamed LLC. | Method of testing immune competency |
-
1987
- 1987-12-17 JP JP31747987A patent/JPH0746103B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH01161151A (en) | 1989-06-23 |
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