ドットブロット、ドットブロット法、ドットブロッティング

関: dot blot、dot blot method

WordNet

travel or traverse (a distance); "This car does 150 miles per hour"; "We did 6 miles on our hike every day"
the syllable naming the first (tonic) note of any major scale in solmization (同)doh, ut
proceed or get along; "How is she doing in her new job?"; "How are you making out in graduate school?"; "Hes come a long way" (同)fare, make_out, come, get along
create or design, often in a certain way; "Do my room in blue"; "I did this piece in wood to express my love for the forest" (同)make
carry on or function; "We could do with a little more help around here" (同)manage
get (something) done; "I did my job" (同)perform
mark with a dot; "dot your `is"
the shorter of the two telegraphic signals used in Morse code (同)dit
scatter or intersperse like dots or studs; "Hills constellated with lights" (同)stud, constellate
make a dot or dots

PrepTutorEJDIC

《疑問文・否定文を作る》 / 《否定命令文を作る》 / 《助動詞とbe動詞のどちらも含まない文に用いて付加疑問を作る》 / 《強意語として》 / 《文》《助動詞とbe動詞のどちらも含まない文に用いて倒置文を作る》 / 《あいづちを打つ場合に》 / 《先行する動詞またはそれを含む述部の代用》 / 《so,nor,neitherで始まる簡略文で》 / …‘を'『する』,行う,果たす / 《通例have done,時にbe doneの形で》…‘を'『終える』,済ませる / …‘を'作る,作り出す / …‘を'『処理する』,整える,片付ける / 〈学課〉‘を'『勉強する』,専攻する / 〈利益・害など〉‘を'与える,もたらす / 〈人〉‘に'『役立つ』,用が足りる(serve) / …の速度で進む,距離を行く / …‘を'見物する / 〈劇〉‘を'上演する;…‘の'役を演じる / …の役目(仕事)をする / 《おもに英》…‘を'だます,かつぐ / 《話》…をへとへとにさせる / 『する』;活動する / 《通例have done,時にbe doneの形で》(…を)『終える』,済ます《+『with』+『名』》 / 〈事が〉『運ぶ』;〈人が〉暮らしていく,健康である / 〈物が〉(…に)『間に合う』,十分である;〈人が〉(…に)役に立つ,向く《+『for』+『名』》 / 《複数形で》すべきこと / 〈C〉《おもに英》大宴会,大パーティー / 〈C〉《英話》詐欺,ぺてん
ド(全音階の第1音)
『点』,ぽち,しみ / (ペンで書いたような)小点;終止符(period) / (通信の)短点 / …‘に'『点を打つ』 / (…を)…‘に'『点在させる』,散在させる《+『名』+『with』+『名』》 / …‘を'打つ,なぐる(hit)
(おもに人称代名詞・固有名詞(人名),thereの後で)had, wouldの短縮形 / (疑問文でwhere,what,whenの後で)didの短縮形;Where'd he go?=Where did he go?
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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2016/06/14 10:50:42」(JST)

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Schematic of the use of two ASO probes on duplicate dot blot filters.

A dot blot (or slot blot) is a technique in molecular biology used to detect biomolecules, and for detecting, analyzing, and identifying proteins. It represents a simplification of the northern blot, Southern blot, or western blot methods. In a dot blot the biomolecules to be detected are not first separated by electrophoresis. Instead, a mixture containing the molecule to be detected is applied directly on a membrane as a dot, and then is spotted through circular templates directly onto the membrane or paper substrate. This differs from the western blot because protein samples are not separated electrophoretically. This is then followed by detection by either nucleotide probes (for a northern blot and southern blot) or antibodies (for a western blot).

The technique offers significant savings in time, as chromatography or gel electrophoresis, and the complex blotting procedures for the gel are not required. However, it offers no information on the size of the target biomolecule. Furthermore, if two molecules of different sizes are detected, they will still appear as a single dot. Dot blots therefore can only confirm the presence or absence of a biomolecule or biomolecules which can be detected by the DNA probes or the antibody.

Because dot blot does not require complicated instrument to operate, lots of dot blot assays are developed using antibodies with high specificity to detect different protein targets. Dot blot is also used to evaluate or screen the effectiveness of the antibodies.

Protocol

Below is just a general guide line for a antigen-antibody-antibody dot blot assay protocol. Specific concentration need to be determined for each assay. Other types of dot blot assays, such as antibody-antigen-antibody can be performed in a similar fashion.

1. spot 1-2 microliter of antigen on to a piece of membrane, let air dry for 30 min or longer;

2. incubate with blocking buffer for 30 min -2 hr;

3. rinse with rinsing buffer, 3x5 min;

4. incubate with primary antibody, 30 min - 2 hr;

5. rinse with rinsing buffer, 3x5 min;

6. incubate with enzyme-labeled secondary antibody, 30 min - 2 hr;

7. rinse with rinsing buffer, 3x5 min;

8. add enzyme substrate, wait 5-10 min;

9. detect by eye or with colorimetric or chemiluminescent imaging system.

Methods

Dot blot is originally performed on a piece of nitrocellulose membrane or PVDF membrane. After the protein samples are spotted onto the membrane, the membrane is placed in a plastic container and sequentially incubated in blocking buffer, antibody solutions, or rinsing buffer on a shaker. Finally, for chemiluminescence imaging, the piece of membrane is wrapped in a transparent plastic film filled with enzyme substrate.

Vacuum-based dot blot apparatus (Bio-dot or Bio-slot from BioRad) has been used to facilitate the rinsing and incubating process by using vacuum to extract the solution from underneath the membrane, which is assembled in between several layers of plates to ensure good seal between sample wells, hold waste solution, and deliver suction force. For chemiluminescence signal detection, the apparatus is dissembled and the membrane is taken out and wrapped in a transparent plastic film filled with enzyme substrate.

Vitrozm's 96 well Zoom Blot method uses an absorption plug to wick away the solution from the membranes, which are individually assembled in each well. The Zoom Plate is a disposable self-standing device, without any external vacuum or set up apparatus, which allows dot blots to be performed much easier and faster. The user never needs to handle a piece of membrane as in older methods. Zoom Blot's 96 well plate format allows convenient signal detection and quantification using 96 well plate reader or imaging system.

What can this test detect?

The sensitive dot blot test can be used to detect the Chlamydia trachomatis infection and other sexually transmitted diseases. Dot blot is used to detect Antidiacyltrehalose Antibodies in Tuberculous patients and Typhoid Fever.

External links

A protocol for the BioRad Dot-Blot Apparatus
Dot Blot using a Zoom Plate

Citations and More Information

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[2]
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[4]
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リンク元	「ドットブロット法」「ドットブロット」「ドットブロッティング」「dot blot method」
関連記事	「do」「blotting」「dot」