大鼠 anti-马 IgG Antibody
ELISA, IHC, WB, FACS, IHC (p), AAb
Polyclonal
IgG
unconjugated
Quick Overview for 大鼠 anti-马 IgG Antibody (ABIN7595375)
抗原
IgG
克隆类型
多克隆
应用范围
ELISA, Immunohistochemistry (IHC), Western Blotting (WB), Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Adaptor Antibody (AAb)
亚型
IgG
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适用
- 马
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宿主
- 大鼠
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原理
- Rat anti horse IgG. Functions as an adapter to convert a primary antibody from one species into another.
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特异性
- Specific for horse IgG
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纯化方法
- Purified
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Goat anti-Horse IgG (F(ab')2 Region) antibody (HRP)
ELISA, IHC, WB Polyclonal IgG HRP
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实验流程
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Western Blot Protocol
- Run the SDS-PAGE at 250V.
- Soak the gel for 15 minutes in transfer buffer, then rinse with DI water.
- Soak the membrane for 5 minutes in DI water.
- Semi-dry transfer at 10V.
- After transfer, rinse the membrane once with PBS, then wash 3× for 5 minutes each with PBS (shaking at 50 RPM).
- Block with 1× PBS containing 1% LFDM for 30 minutes at room temperature (shake at 50 RPM).
- Rinse with 70 ml of PBS.
- Add your primary antibodies and incubate overnight in the fridge (3–5°C).
- Wash the membrane 5 times (5 minutes each) with PBST containing 0.1% Tween-20 (use 100 ml, shake at 50 RPM).
- Add the Adaptor-Secondary of your choice at 1:5000 dilution and incubate with shake for 30 minutes.
- Dip in DI water (or 1x PBS).
- Incubate with your secondary antibody for example, HRP-Rb anti-Ch (diluted 1:20,000). Use 5 ml per membrane (6×8 cm). Incubate for 30 minutes to 1 hour at room temperature, shaking at 50 RPM.
- Wash 4 times (5 minutes each) with 100 ml of PBST, shaking at 50 RPM.
- Wash 6 more times with 100 ml of PBS, shaking at 50 RPM.
- Add ECL substrate.
- Scan the blot on a C-Digit at high resolution (12 minutes).
- Cut the issue at 5 µm
- Drain and Dry
- Dewax and Dehydrate
- HIER using Thermo's PT Module - HIER buffer: Tris-EDTA pH 9.00
- Stain the slides: Buffer wash - Peroxide Block (5 min) - Buffer wash (2x) - Protein Block (5 min) - Buffer wash (2x) - Primary antibody (30 min) - Buffer wash (2x) - Adaptor-Secondary - Buffer wash - HRP-Polymer (30 min) - Buffer wash (3x) - Water wash (1x)
- Add DAB
- Counterstain
- Coverslip
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限制
- 仅限研究用
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状态
- Lyophilized
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溶解方式
- Reconstitute in 500 µL DI water.
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缓冲液
- Contains Signal Enhancer and Background Quencher. Preservative: 0.02% benzalkonium chloride
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储存液
- Benzalkonium chloride
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储存条件
- 4 °C,-20 °C
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储存方法
- Maintain the lyophilised/reconstituted antibodies frozen at -20°C for long term storage and refrigerated at 2-8°C for a shorter term.
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有效期
- 12 months
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- IgG
抗原
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