HA單株抗體 -瓊脂糖(agarose) | Abbkine貨號ABT2043
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HA單株抗體 -瓊脂糖(agarose) | Abbkine貨號ABT2043
Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6)
Abbkine提供一系列高度特異性單克隆和多克隆抗體，tag antibody抗體系列商品如 His tag、GST tag、S tag、HA tag、DDDDK tag、Myc tag、MBP tag和 mCherry tag等單/多株Tag抗體。另具生物素(Biotin)、HRP、AbFluor™ 染料、瓊脂糖(agarose)和磁珠偶聯物(magnetic beads)標誌。人類流感血凝素 (Human influenza hemagglutinin , HA) 是人類病毒感染所必需的表面糖蛋白(surface glycoprotein)。 HA tag源自氨基酸98-106的HA分子(HA-molecule)，已被廣泛用作表達載體中的通用表位標籤。許多重組蛋白已被改造以表達 HA 標籤，而 HA 標籤似乎不會干擾重組蛋白的生物活性或生物分佈。
• 結合容量：1 mg 抗體與 1 mL 填充瓊脂糖偶聯，每 mL 產品含有 1 mL 瓊脂糖和 1 mL 儲存緩衝液。容量：≥0.6 mg HA-Tag蛋白/mL瓊脂糖 • 成分：儲存緩衝液：50% 凝膠漿懸浮於 PBS 中，pH 7.4，含 0.02% 疊氮化鈉。Beads Size: 45-165 μm
His Tag單株抗體(HRP) | Flag-tag單株抗體(HRP) | GFP單株抗體(HRP) | GST單株抗體(HRP) | HA單株抗體(HRP)
A. Preparation of Agarose
Note: Per 500 μL of protein sample add 20 μL (total 40 μL suspension) Agarose.
(1) Add Agarose to a 1.5 mL centrifuge tube. Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.
(2) Add 1 mL 1×TBS to re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3times. Add20 μL1×TBS to re-suspend Agarose.
B. Immunoprecipitation
(1) Add 500 μL protein samples to the processed Agarose, and incubate at room temperature for 1-2 h or overnight at 4℃(It isrecommended to use vertical rotating mixer with Low-speed rotation).
(2) Centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant.
(3) Add 1mL 1×TBS, and re-suspend Agarose, centrifuge at 800 rpm for 2 min at 4℃, remove the supernatant, repeat 3-5times,until OD280 of the supernatant is lesser than 0.05.
(4) Elution
a）Denatured elution: This method is suitable for SDS-PAGE and Western Blotting analysis of elution samples. Add100μL(5
times volume of Agarose) 1×SDS-PAGE Loading Buffer to the tube and mix well, incubate at 100℃for 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
analysis. Add 100 μL (5 times volume of Agarose) HA Peptide (0.1-0.2 μg/mL) to the tube and mix well, incubate at 4℃for 1-2h(It is recommended to use vertical rotating mixer with Low-speed rotation), then centrifuge at 800 rpm for 2 min at 4℃, 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℃/-80℃forlong-term. It is recommended to add 100 μL 1×SDS-PAGE Loading Buffer to Agarose precipitation to test theeffectofimmunoprecipitation and elution.
b）Acid elution: This method can maintain their original biological activity, elution can be used for functional analysis. Add100μL(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℃, and collect thesupernatantwhich is HA-Tag protein and its complex to a new tube, and immediately add 10 μL 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℃/-80℃ for long-term
It is recommended to add 100 μL1×SDS-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 μL Acid Elution Buffer (0.1 MGlycine-HCl, pH2.8) and 15 μL Neutralizing Buffer (1 M Tris-HCl, pH 8.5).
| Cat.No | Name | Size1 |
| KTD104-EN | Universal IP/Co-IP Toolkit (Magnetic Beads) | 20T |
| KTD105-EN | Universal IP/Co-IP Toolkit (Agarose) | 20T |
| A25012 | IPKine™ HRP, Goat Anti-Mouse IgG LCS | 100 μL |
| A25022 | IPKine™ HRP, Mouse Anti-Rabbit IgG LCS | 100 μL |
| A25112 | IPKine™ HRP, Goat Anti-Mouse IgG HCS | 100 μL |
| A25122 | IPKine™ HRP, Mouse Anti-Rabbit IgG HCS | 100 μL |
| A25222 | IPKine™ HRP, Goat Anti-Rabbit IgG HCS | 100 μL |
| KTI2014 | IPKine™ Anti-DDDDK Magnetic IP Kit | 20T |
| KTI2024 | IPKine™ Anti-GFP Magnetic IP Kit | 20T |
| KTI2034 | IPKine™ Anti-GST Magnetic IP Kit | 20T |
| KTI2044 | IPKine™ Anti-HA Magnetic IP Kit | 20T |
| KTI2064 | IPKine™ Anti-Myc Magnetic IP Kit | 20T |
| ABT2013 | Agarose Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) | 1 mL |
| ABT2014 | Magnetic Beads Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) | 1 mL |
| ABT2024 | Magnetic Beads Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3) | 1 mL |
| ABT2023 | Agarose Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3) | 1 mL |
| ABT2043 | Agarose Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6) | 1 mL |
| ABT2044 | Magnetic Beads Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6) | 1 mL |
| ABT2053 | Agarose Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3) | 1 mL |
| ABT2054 | Magnetic Beads Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3) | 1 mL |
| ABT2063 | Agarose Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5) | 1 mL |
| ABT2064 | Magnetic Beads Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5) | 1 mL |
| ABT2173 | Agarose Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5) | 1 mL |
| ABT2174 | Magnetic Beads Conjugated Anti-V5 Tag Mouse Monoclonal Antibody (11D5) | 1 mL |
| ABT2010 | Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) | 50 μL |
| ABT2011 | Anti-DDDDK Tag Rabbit Polyclonal Antibody | 50 μL |
| ABT2015 | HRP Conjugated Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) | 50 μL |
| ABT2020 | Anti-GFP Tag Mouse Monoclonal Antibody (3D3) | 50 μL |
| ABT2025 | HRP Conjugated Anti-GFP Tag Mouse Monoclonal Antibody (3D3) | 50 μL |
| ABT2021 | Anti-GFP Tag Rabbit Polyclonal Antibody | 50 μL |
| ABT2030 | Anti-GST Tag Mouse Monoclonal Antibody (2A8) | 50 μL |
| ABT2035 | HRP Conjugated Anti-GST Tag Mouse Monoclonal Antibody (2A8) | 50 μL |
| ABT2040 | Anti-HA Tag Mouse Monoclonal Antibody (4F6) | 50 μL |
| ABT2041 | Anti-HA Tag Rabbit Polyclonal Antibody | 50 μL |
| ABT2045 | HRP Conjugated Anti-HA Tag Mouse Monoclonal Antibody (4F6) | 50 μL |
| ABT2050 | Anti-His Tag Mouse Monoclonal Antibody (5C3) | 50 μL |
| ABT2051 | Anti-His Tag Rabbit Polyclonal Antibody | 50 μL |
| ABT2055 | HRP Conjugated Anti-His Tag Mouse Monoclonal Antibody (5C3) | 50 μL |
| ABT2060 | Anti-Myc Tag Mouse Monoclonal Antibody (2D5) | 50 μL |
| ABT2061 | Anti-Myc Tag Rabbit Polyclonal Antibody | 50 μL |
| ABT2065 | HRP Conjugated Anti-Myc Tag Mouse Monoclonal Antibody (2D5) | 50 μL |
| ABT2170 | Anti-V5 Tag Mouse Monoclonal Antibody (11D5) | 50 μL |
| ABT2070 | Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) | 50 μL |
| ABT2080 | Anti-mCherry Tag Mouse Monoclonal Antibody (9D3) | 50 μL |
| ABT2090 | Anti-E2 Tag Mouse Monoclonal Antibody (12T4) | 50 μL |
| ABT2100 | Anti-HSV Tag Mouse Monoclonal Antibody (16T2) | 50 μL |
| ABT2110 | Anti-KT3 Tag Mouse Monoclonal Antibody (14D8) | 50 μL |
| ABT2120 | Anti-RFP Tag Mouse Monoclonal Antibody (9D1) | 50 μL |
| ABT2130 | Anti-S Tag Mouse Monoclonal Antibody (9T10) | 50 μL |
| ABT2150 | Anti-T7 Tag Mouse Monoclonal Antibody (6D4) | 50 μL |
| ABT2160 | Anti-Trx Tag Mouse Monoclonal Antibody (14D4) | 50 μL |
| ABT2180 | Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) | 50 μL |
| ABT2190 | Anti-CBP Tag Monoclonal Antibody (12H5) | 50 μL |
| ABT2200 | Anti-TAP Tag Monoclonal Antibody (4H2) | 50 μL |
| ABT2210 | Anti-Avi-Tag Monoclonal Antibody (5G11) | 50 μL |
| ABT2220 | Anti-SRT-Tag Monoclonal Antibody (11G3) | 50 μL |
| ABT2230 | Anti-Strep-Tag II Monoclonal Antibody (8C12) | 50 μL |
| ABT2240 | Anti-mStrawberry Monoclonal Antibody (4C9) | 50 μL |
| ABT2250 | Anti-EYFP-Tag Monoclonal Antibody (10T3) | 50 μL |
| ABT2260 | Anti-mOrange Monoclonal Antibody (9A10) | 50 μL |
| ABT2270 | Anti-AmCyan Monoclonal Antibody (8T2) | 50 μL |
| ABT2280 | Anti-ECFP-Tag Monoclonal Antibody (6B11) | 50 μL |
| ABT2290 | Anti-EBFP Monoclonal Antibody (8B5) | 50 μL |
| ABT2300 | Anti-Nano-Tag9 Monoclonal Antibody (11T3) | 50 μL |