亚洲av无码无在线观看红杏,かしこまりました中文在线,国产精品免费一区二区三区视频,老同学3免费完整版在线观看

您好,歡迎進(jìn)入上海力敏實業(yè)有限公司網(wǎng)站!
一鍵分享網(wǎng)站到:
您現(xiàn)在的位置:首頁 >> 產(chǎn)品中心 >> 生化試劑、抗體、血清 >> 常規(guī)生化試劑 >> B-11335VECTOR EEL生物素大葉黃楊Europaeus凝集素

VECTOR EEL生物素大葉黃楊Europaeus凝集素

  • 更新時間:  2023-07-24
  • 產(chǎn)品型號:  B-11335
  • 簡單描述
  • VECTOR EEL生物素大葉黃楊Europaeus凝集素
    Detection of Glycoproteins using Lectins in Histochemistry,
    ELISA, and Western Blot Applications
詳細(xì)介紹

 

VECTOR EEL生物素大葉黃楊Europaeus凝集素

The following protocols offer guidelines for assay development using lectin-based detection of glycoproteins
present in tissue sections, adsorbed onto microtiter plates, or transferred from electrophoretic
gels onto nitrocellulose or PVDF membranes.
Histochemistry:
1a. Staining procedure for paraffin sections: Deparaffinize and hydrate tissue sections through
xylenes or other clearing agents and graded alcohol series and rinse for 5 minutes in tap water.
If required, retrieve antigens using the Antigen Unmasking Solution (H-3300 or H-3301).
1b. Staining procedure for frozen sections: Air dry sections. Immediay before staining, fix
sections with acetone. Transfer slices to buffer. If endogenous enzyme activities are present,
inactivate using appropriate methods.
2. Perform Streptavidin/Biotin blocking if required following kit instructions (SP-2002). Do not
use SP-2001. Block non-specific binding by incubating section with Carbo-Free™ Blocking
Solution (Cat. No. SP-5040) for 30 minutes at room temperature. Blot excess blocking solution
from the sections.
3. Apply biotinylated lectin at approximay 2-20 μg/ml in PBS (10 mM sodium phosphate, 150
mM NaCl, pH 7.4) to the sections and incubate for 30 minutes at room temperature. Wash with
TPBS (PBS + 0.05% Tween™20).
4. Prepare VECTASTAIN®
® ABC (peroxidase, Cat. No. PK-6100) or VECTASTAIN® ABC-AP
(alkaline phosphatase, Cat. No. AK-5000) reagents according to the kit instructions. Apply to the
sections and incubate for 30 minutes at room temperature. Wash with TPBS.
5. Apply an appropriate precipitating substrate for the enzyme system used in step 4. For peroxidase,
ImmPACT™ DAB (Cat. No. SK-4105) is recommended; for alkaline phosphatase, Vector® Red
(Cat. No. SK-5100). Rinse in tap water.
6. Counterstain (optional), clear and mount. For galactose or GalNAc-specific lectins avoid mounting
in glycerol-based mounting media.
ELISA:
1. Adsorb target protein to microtiter plate by placing 50-200 μl of approximay 3 μg/ml glycoprotein
solution into the desired wells. Some wells may be left untreated as negative controls. Incubate at
37 oC for 1 hour. Wash wells three times with TPBS (PBS + 0.05% Tween™20).
2. Block non-specific binding by filling each well to the brim with Carbo-Free™ Blocking Solution
(Cat. No. SP-5040) for 30 minutes at room temperature. Wash wells three times with TPBS.
3. Apply 50-200 μl of approximay 2-20 μg/ml biotinylated lectin in PBS to the wells and incubate
for 30 minutes at room temperature. Wash wells three times with TPBS.

4. Prepare VECTASTAIN®
® ABC (peroxidase, Cat. No. PK-6100) or VECTASTAIN® ABC-AP
(alkaline phosphatase, Cat. No. AK-5000) reagents according to the kit instructions. Apply to the
wells and incubate for 30 minutes at room temperature. Wash wells three times with TPBS.
5. Apply an appropriate non-precipitating substrate for the enzyme system used in step 4. For
peroxidase, ABTS (Cat. No. SK-4500) is recommended; for alkaline phophatase, pNPP (Cat. No.
SK-5900).
6. Quantify the colored reaction product by spectrophotometry.
Western Blot:
1. Perform electrophoresis and transfer proteins to a membrane according to standard procedures.
2. Block non-specific binding by incubating the membrane in Carbo-Free™ Blocking Solution (Cat.
No. SP-5040) for 30 minutes at room temperature. Use a sufficient volume to compley cover
the membrane.
3. Incubate membrane in PBS containing approximay 2-20 μg/ml biotinylated lectin for 30 minutes
at room temperature. Wash with TPBS (PBS +0.05% Tween™20).
4. Prepare VECTASTAIN®
® ABC (peroxidase, Cat. No. PK-6100) or VECTASTAIN® ABC-AP
(alkaline phosphatase, Cat. No. AK-5000) reagents according to the kit instructions. Incubate the
membrane in the reagent for 30 minutes at room temperature. Wash with TPBS.
5. Apply an appropriate substrate for the enzyme system used in step 4. For peroxidase, DuoLuX™
Chemiluminescent/Fluorescent Substrate for Peroxidase (Cat. No. SK-6604) or ImmPACT™ DAB
(Cat. No. SK-4105) are recommended; for alkaline phosphatase, Chemiluminescent/Fluorescent
Substrate for Alkaline Phosphatase (Cat. No. SK-6605) or BCIP/NBT (Cat. No. SK-5400) are
recommended.
VECTOR EEL生物素大葉黃楊Europaeus凝集素

Negative Controls
Negative controls should be run in parallel in each of the above described methodologies to validate
binding results. When applying lectins, one of the most appropriate negative controls is to preabsorb
the lectin with a concentration of a defined sugar, with which, the lectin has a known high affinity.
Vector Labs offers a series of sugars that are intended for such a purpose.
The lectin is diluted to a suitable working concentration in a solution containing approximay
200 mM to 500 mM of the sugar. This mixture is left to bind at room temperature for 30 to 60 min.
Following this absorption incubation, the mixture is substituted into the procedure in place of the unabsorbed
lectin and incubated under the same conditions. The subsequent detection procedure is followed
as for the test method. In most cases the vast majority of lectin binding to the tissue section (membrane
blot, etc.) will be eliminated. Some trace binding to the section (blot etc) may still be present under
these conditions and probably indicates presence of secondary or tertiary sugar preferences. These negative
control results should be compared with the test results to determine specificity of binding

 

詳細(xì)產(chǎn)品信息可和選購


留言框

  • 產(chǎn)品:

  • 您的單位:

  • 您的姓名:

  • 聯(lián)系電話:

  • 常用郵箱:

  • 省份:

  • 詳細(xì)地址:

  • 補充說明:

  • 驗證碼:

    請輸入計算結(jié)果(填寫阿拉伯?dāng)?shù)字),如:三加四=7
久久亚洲国产成人影院| 一本一道AV无码中文字幕| 黑人巨大精品欧美黑寡妇| 亚洲AV无码不卡久久| 色翁荡熄又大又硬又粗又视频| 亚洲线精品一区二区三区影音先锋| 被强行灌满精子的少妇| 青春草在线视频观看| 国语自产少妇精品视频| 男人扒开添女人下部免费视频| 色窝窝亚洲av网在线观看| 国产精品爱久久久久久久| 亚洲午夜无码久久| 色婷婷综合久久久久中文一区二区 | 精品亚洲成A人片在线观看少妇| 久久久久久曰本av免费免费| 亚洲国产精品久久无码中文字蜜桃 | 亚洲av无码一区二区三区人| 国产麻豆成人精品av| 国产免费AV片无码永久免费| 免费看少妇高潮成人片| 日韩精品一区二区三区中文| 国产精品毛片无码一区二区蜜桃| 巨胸喷奶水WWW视频网站| 无码av天天av天天爽| 欧美人妻一区黄a片| 永久免费a∨片在线观看| 无码精油按摩潮喷在播放| 偿还HD韩国中文版| 国产又色又爽无遮挡免费| 亚洲va韩国va欧美va| 国精产品一区二区三区公司| 国产福利美女福利视频免费看| 亚洲av无码一区二区乱子伦| 精品亚洲成a人无码成a在线观看| AV无码精品久久久久精品免费| 国产福利一区二区三区在线观看| 中国人在线观看免费高清| 亚洲熟妇丰满xxxxx国语| 国产精品无码A∨精品影院| 日本精品人妻无码免费大全|