您好,歡迎進入上海力敏實業有限公司網站!
一鍵分享網站到:
您現在的位置:首頁 >> 產品中心 >> 生化試劑、抗體、血清 >> 常規生化試劑 >> VECTOR MAL B-1265懷槐凝集素Biotinylated Maackia Amurensis Lectin

懷槐凝集素Biotinylated Maackia Amurensis Lectin

  • 更新時間:  2023-07-25
  • 產品型號:  VECTOR MAL B-1265
  • 簡單描述
  • 懷槐凝集素Biotinylated Maackia Amurensis Lectin
    Biotinylated Maackia Amurensis Lectin (MAL) II
    Detection of Glycoproteins using Lectins in Histochemistry,
    ELISA, and Western Blot Applications
詳細介紹

懷槐凝集素Biotinylated Maackia Amurensis Lectin

Detection of Glycoproteins using Lectins in Histochemistry,
ELISA, and Western Blot Applications
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.
Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA 94010 U.S.A.
 

懷槐凝集素Biotinylated Maackia Amurensis Lectin

VECTOR
LABORATORIES
VECTOR
LABORATORIES
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.
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

 

詳細產品信息可和選購


留言框

  • 產品:

  • 您的單位:

  • 您的姓名:

  • 聯系電話:

  • 常用郵箱:

  • 省份:

  • 詳細地址:

  • 補充說明:

  • 驗證碼:

    請輸入計算結果(填寫阿拉伯數字),如:三加四=7
友情鏈接: 等離子去膠機       電液伺服萬能材料試驗機       電液伺服液壓萬能試驗機       電子拉伸萬能試驗機       材料拉伸試驗機       金剛石鉆孔機       北斗定位器       控溫式遠紅外消煮爐       切膠儀       無塵紙       金屬液氮冷凍箱       傳力接頭       磁座鉆       LEM電壓傳感器       鋁儲罐      
主站蜘蛛池模板: 亚洲国产一区二区三区在线观看| 嗯!啊!使劲用力在线观看| AV无码精品一区二区三区宅噜噜 | 男女一进一出呻吟的动态图| 国产一级一级一级国产片| 国产a免费观看| 国产精品亚洲精品日韩动图| 97国产免费全部免费观看| 女大学生沙龙室3| 中国一级毛片视频免费看| 日本不卡免费新一二三区| 久久综合丝袜日本网| 欧美一区二区三区精品影视 | 久久伊人成人网| 最近中文字幕mv免费高清电影| 亚洲小说区图片区另类春色| 狠狠人妻久久久久久综合蜜桃| 六月丁香综合网| 美女扒开尿口让男人桶免费网站| 国产丫丫视频私人影院| 韩国理论福利片午夜| 国产成人亚综合91精品首页| 亚洲最大成人网色香蕉| 国产精品无码V在线观看| 91免费视频网| 国内精品久久久久影视| 99精品国产高清一区二区麻豆| 好男人社区在线www| 一级毛片在播放免费| 成人精品一区二区三区中文字幕| 久久久一本精品99久久精品66| 日本肉动漫无遮挡无删减在线观看 | 亚洲熟妇AV乱码在线观看| 激情综合婷婷色五月蜜桃| 你是我的女人中文字幕高清| 穿透明白衬衫喷奶水在线播放| 冲田杏梨在线精品二区| 美国式禁忌4桥矿超棒| 嘟嘟嘟www在线观看免费高清| 色综合久久天天综合| 抬头见喜全集免费版|