{"id":53037,"date":"2025-10-09T17:01:13","date_gmt":"2025-10-09T09:01:13","guid":{"rendered":"https:\/\/biopioneer.com.tw\/?p=53037"},"modified":"2025-11-03T13:44:10","modified_gmt":"2025-11-03T05:44:10","slug":"ha%e5%96%ae%e6%a0%aa%e6%8a%97%e9%ab%94-%e7%93%8a%e8%84%82%e7%b3%96agarose-abbkine%e8%b2%a8%e8%99%9fabt2043","status":"publish","type":"post","link":"https:\/\/biopioneer.com.tw\/?p=53037","title":{"rendered":"HA\u55ae\u682a\u6297\u9ad4 -\u74ca\u8102\u7cd6(agarose)\u00a0| Abbkine\u8ca8\u865fABT2043"},"content":{"rendered":"

HA\u55ae\u682a\u6297\u9ad4 -\u74ca\u8102\u7cd6(agarose)\u00a0| <\/span><\/strong>Abbkine\u8ca8\u865f<\/strong>ABT2043<\/span><\/span><\/a><\/span><\/p>\n

Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)<\/p>\n

Abbkine\u63d0\u4f9b\u4e00\u7cfb\u5217\u9ad8\u5ea6\u7279\u7570\u6027\u55ae\u514b\u9686\u548c\u591a\u514b\u9686\u6297\u9ad4\uff0ctag antibody\u6297\u9ad4\u7cfb\u5217\u5546\u54c1\u5982 His tag\u3001GST tag\u3001S tag\u3001HA tag\u3001DDDDK tag\u3001Myc tag\u3001MBP tag\u548c mCherry tag\u7b49\u55ae\/\u591a\u682aTag\u6297\u9ad4\u3002\u53e6\u5177\u751f\u7269\u7d20(Biotin)\u3001HRP\u3001AbFluor\u2122 \u67d3\u6599\u3001\u74ca\u8102\u7cd6(agarose)\u548c\u78c1\u73e0\u5076\u806f\u7269(magnetic beads)\u6a19\u8a8c\u3002\u4eba\u985e\u6d41\u611f\u8840\u51dd\u7d20 (Human influenza hemagglutinin , HA) \u662f\u4eba\u985e\u75c5\u6bd2\u611f\u67d3\u6240\u5fc5\u9700\u7684\u8868\u9762\u7cd6\u86cb\u767d(surface glycoprotein)\u3002 HA tag\u6e90\u81ea\u6c28\u57fa\u917898-106\u7684HA\u5206\u5b50(HA-molecule)\uff0c\u5df2\u88ab\u5ee3\u6cdb\u7528\u4f5c\u8868\u9054\u8f09\u9ad4\u4e2d\u7684\u901a\u7528\u8868\u4f4d\u6a19\u7c64\u3002\u8a31\u591a\u91cd\u7d44\u86cb\u767d\u5df2\u88ab\u6539\u9020\u4ee5\u8868\u9054 HA \u6a19\u7c64\uff0c\u800c HA \u6a19\u7c64\u4f3c\u4e4e\u4e0d\u6703\u5e72\u64fe\u91cd\u7d44\u86cb\u767d\u7684\u751f\u7269\u6d3b\u6027\u6216\u751f\u7269\u5206\u4f48\u3002<\/p>\n

\u2022 \u7d50\u5408\u5bb9\u91cf\uff1a1 mg \u6297\u9ad4\u8207 1 mL \u586b\u5145\u74ca\u8102\u7cd6\u5076\u806f\uff0c\u6bcf mL \u7522\u54c1\u542b\u6709 1 mL \u74ca\u8102\u7cd6\u548c 1 mL \u5132\u5b58\u7de9\u885d\u6db2\u3002\u5bb9\u91cf\uff1a\u22650.6 mg HA-Tag\u86cb\u767d\/mL\u74ca\u8102\u7cd6<\/span>\r\n\u2022 \u6210\u5206\uff1a\u5132\u5b58\u7de9\u885d\u6db2\uff1a50% \u51dd\u81a0\u6f3f\u61f8\u6d6e\u65bc PBS \u4e2d\uff0cpH 7.4\uff0c\u542b 0.02% \u758a\u6c2e\u5316\u9209\u3002Beads Size: 45-165 \u03bcm\r\n\r\n<\/span><\/pre>\n
<\/p>\n
His Tag\u55ae\u682a\u6297\u9ad4(HRP)\u00a0<\/strong><\/span><\/a>\u00a0|\u00a0<\/a>Flag-tag\u55ae\u682a\u6297\u9ad4(HRP<\/strong>)<\/strong><\/a>\u00a0|\u00a0<\/a>GFP\u55ae\u682a\u6297\u9ad4<\/a>(HRP) |\u00a0<\/a>GST\u55ae\u682a\u6297\u9ad4<\/a>(HRP) |\u00a0<\/a>HA\u55ae\u682a\u6297\u9ad4<\/a>(HRP)<\/a><\/span><\/p>\n
A. Preparation of Agarose
\nNote: Per 500 \u03bcL of protein sample add 20 \u03bcL (total 40 \u03bcL suspension) Agarose.<\/p>\n
(1) Add Agarose to a 1.5 mL centrifuge tube. Centrifuge at 800 rpm for 2 min at 4\u2103, remove the supernatant.
\n(2) Add 1 mL 1\u00d7TBS to re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4\u2103, remove the supernatant, repeat 3times. Add20 \u03bcL1\u00d7TBS to re-suspend Agarose.
\nB. Immunoprecipitation
\n(1) Add 500 \u03bcL protein samples to the processed Agarose, and incubate at room temperature for 1-2 h or overnight at 4\u2103(It isrecommended to use vertical rotating mixer with Low-speed rotation).
\n(2) Centrifuge at 800 rpm for 2 min at 4\u2103, remove the supernatant.<\/p>\n
(3) Add 1mL 1\u00d7TBS, and re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4\u2103, remove the supernatant, repeat 3-5times,until OD280 of the supernatant is lesser than 0.05.<\/p>\n
(4) Elution
\na\uff09Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add100\u03bcL(5
\ntimes volume of Agarose) 1\u00d7SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100\u2103for 5 min, thenCentrifugeat800 rpm for 1 min, and collect the supernatant to a new tube for SDS-PAGE and Western Blotting analysis. Competitive elution of peptide: This method can maintain their original biological activity, elution can be used for functional
\nanalysis. Add 100 \u03bcL (5 times volume of Agarose) HA Peptide (0.1-0.2 \u03bcg\/mL) to the tube and mix well, incubate at 4\u2103for 1-2h(It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4\u2103, andcollectthe supernatant which is HA-Tag protein and its complex to a new tube. In order to improve the elution efficiency, theincubationtime can be Increased or repeat elution. Place HA-Tag protein and its complex on ice to be used, or store at -20\u2103\/-80\u2103forlong-term. It is recommended to add 100 \u03bcL 1\u00d7SDS-PAGE Loading Buffer to Agarose precipitation to test theeffectofimmunoprecipitation and elution.<\/p>\n
b\uff09Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add100\u03bcL(5 times volume of Agarose) Elution Buffer to the tube and mix well, incubate at room temperature for 5-10 min (It is recommendedto use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4\u2103, and collect thesupernatantwhich is HA-Tag protein and its complex to a new tube, and immediately add 10 \u03bcL Neutralization Buffer to adjust thepHto7.0-8.0. In order to improve the elution efficiency, elution can be repeated, and combine the same samples. Place HA-Tagproteinand its complex on ice to be used, or store at -20\u2103\/-80\u2103 for long-term<\/p>\n
It is recommended to add 100 \u03bcL1\u00d7SDS-PAGELoading Buffer to Agarose precipitation to test the effect of immunoprecipitation and elution. Note: a) For a few samples, due to differences in target proteins, the binding of HA-Tag and Anti-HAantibodyisverystrong, and the effect of Acid elution and Competitive elution of peptide may be poor. Therefore, SDS-PAGELoadingBuffer denaturation elution method is recommended as a priority; b) Due to the difference of target protein, theelutionefficiency of acid elution method also varies to some extent. If the requirement of elution efficiency is high, thepHvalueof acidic eluent can be adjusted appropriately between 2.5-3.1, and the pH value or quantity of correspondingneutralizing solution should be adjusted appropriately. For example, 100 \u03bcL Acid Elution Buffer (0.1 MGlycine-HCl, pH2.8) and 15 \u03bcL Neutralizing Buffer (1 M Tris-HCl, pH 8.5).<\/p>\n
\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Cat.No<\/td>\nName<\/td>\nSize1<\/td>\n<\/tr>\n
KTD104-EN<\/td>\nUniversal IP\/Co-IP Toolkit (Magnetic Beads)<\/td>\n20T<\/td>\n<\/tr>\n
KTD105-EN<\/td>\nUniversal IP\/Co-IP Toolkit (Agarose)<\/td>\n20T<\/td>\n<\/tr>\n
A25012<\/td>\nIPKine\u2122 HRP, Goat Anti-Mouse IgG LCS<\/td>\n100 \u03bcL<\/td>\n<\/tr>\n
A25022<\/td>\nIPKine\u2122 HRP, Mouse Anti-Rabbit IgG LCS<\/td>\n100 \u03bcL<\/td>\n<\/tr>\n
A25112<\/td>\nIPKine\u2122 HRP, Goat Anti-Mouse IgG HCS<\/td>\n100 \u03bcL<\/td>\n<\/tr>\n
A25122<\/td>\nIPKine\u2122 HRP, Mouse Anti-Rabbit IgG HCS<\/td>\n100 \u03bcL<\/td>\n<\/tr>\n
A25222<\/td>\nIPKine\u2122 HRP, Goat Anti-Rabbit IgG HCS<\/td>\n100 \u03bcL<\/td>\n<\/tr>\n
KTI2014<\/td>\nIPKine\u2122 Anti-DDDDK Magnetic IP Kit<\/td>\n20T<\/td>\n<\/tr>\n
KTI2024<\/td>\nIPKine\u2122 Anti-GFP Magnetic IP Kit<\/td>\n20T<\/td>\n<\/tr>\n
KTI2034<\/td>\nIPKine\u2122 Anti-GST Magnetic IP Kit<\/td>\n20T<\/td>\n<\/tr>\n
KTI2044<\/td>\nIPKine\u2122 Anti-HA Magnetic IP Kit<\/td>\n20T<\/td>\n<\/tr>\n
KTI2064<\/td>\nIPKine\u2122 Anti-Myc Magnetic IP Kit<\/td>\n20T<\/td>\n<\/tr>\n
ABT2013<\/td>\nAgarose Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2014<\/td>\nMagnetic Beads Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2024<\/td>\nMagnetic Beads Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2023<\/td>\nAgarose Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2043<\/td>\nAgarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2044<\/td>\nMagnetic Beads Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2053<\/td>\nAgarose Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2054<\/td>\nMagnetic Beads Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2063<\/td>\nAgarose Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2064<\/td>\nMagnetic Beads Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2173<\/td>\nAgarose Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2174<\/td>\nMagnetic Beads Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5)<\/td>\n1 mL<\/td>\n<\/tr>\n
ABT2010<\/td>\nAnti-DDDDK Tag Mouse Monoclonal Antibody (1B10)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2011<\/td>\nAnti-DDDDK Tag Rabbit Polyclonal Antibody<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2015<\/td>\nHRP Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2020<\/td>\nAnti-GFP Tag Mouse Monoclonal Antibody (3D3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2025<\/td>\nHRP Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2021<\/td>\nAnti-GFP Tag Rabbit Polyclonal Antibody<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2030<\/td>\nAnti-GST Tag Mouse Monoclonal Antibody (2A8)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2035<\/td>\nHRP Conjugated Anti-GST Tag Mouse Monoclonal Antibody (2A8)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2040<\/td>\nAnti-HA Tag Mouse Monoclonal Antibody (4F6)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2041<\/td>\nAnti-HA Tag Rabbit Polyclonal Antibody<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2045<\/td>\nHRP Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2050<\/td>\nAnti-His Tag Mouse Monoclonal Antibody (5C3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2051<\/td>\nAnti-His Tag Rabbit Polyclonal Antibody<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2055<\/td>\nHRP Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2060<\/td>\nAnti-Myc Tag Mouse Monoclonal Antibody (2D5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2061<\/td>\nAnti-Myc Tag Rabbit Polyclonal Antibody<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2065<\/td>\nHRP Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2170<\/td>\nAnti-V5 Tag Mouse Monoclonal Antibody (11D5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2070<\/td>\nAnti-MBP Tag Mouse Monoclonal Antibody (9Y5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2080<\/td>\nAnti-mCherry Tag Mouse Monoclonal Antibody (9D3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2090<\/td>\nAnti-E2 Tag Mouse Monoclonal Antibody (12T4)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2100<\/td>\nAnti-HSV Tag Mouse Monoclonal Antibody (16T2)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2110<\/td>\nAnti-KT3 Tag Mouse Monoclonal Antibody (14D8)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2120<\/td>\nAnti-RFP Tag Mouse Monoclonal Antibody (9D1)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2130<\/td>\nAnti-S Tag Mouse Monoclonal Antibody (9T10)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2150<\/td>\nAnti-T7 Tag Mouse Monoclonal Antibody (6D4)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2160<\/td>\nAnti-Trx Tag Mouse Monoclonal Antibody (14D4)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2180<\/td>\nAnti-VSV-G-Tag Mouse Monoclonal Antibody (14D2)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2190<\/td>\nAnti-CBP Tag Monoclonal Antibody (12H5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2200<\/td>\nAnti-TAP Tag Monoclonal Antibody (4H2)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2210<\/td>\nAnti-Avi-Tag Monoclonal Antibody (5G11)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2220<\/td>\nAnti-SRT-Tag Monoclonal Antibody (11G3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2230<\/td>\nAnti-Strep-Tag II Monoclonal Antibody (8C12)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2240<\/td>\nAnti-mStrawberry Monoclonal Antibody (4C9)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2250<\/td>\nAnti-EYFP-Tag Monoclonal Antibody (10T3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2260<\/td>\nAnti-mOrange Monoclonal Antibody (9A10)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2270<\/td>\nAnti-AmCyan Monoclonal Antibody (8T2)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2280<\/td>\nAnti-ECFP-Tag Monoclonal Antibody (6B11)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2290<\/td>\nAnti-EBFP Monoclonal Antibody (8B5)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n
ABT2300<\/td>\nAnti-Nano-Tag9 Monoclonal Antibody (11T3)<\/td>\n50 \u03bcL<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
 <\/p>\n","protected":false},"excerpt":{"rendered":"
HA\u55ae\u682a\u6297\u9ad4 -\u74ca\u8102\u7cd6(agarose)\u00a0| Abbkine\u8ca8\u865fABT2043 Agarose Conjuga […]<\/p>\n","protected":false},"author":3,"featured_media":29551,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5262],"tags":[7001],"_links":{"self":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/53037"}],"collection":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=53037"}],"version-history":[{"count":2,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/53037\/revisions"}],"predecessor-version":[{"id":53051,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/53037\/revisions\/53051"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/29551"}],"wp:attachment":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=53037"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=53037"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=53037"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}