豚鼠 anti-兔 IgG (Heavy & Light Chain) Antibody - Preadsorbed
(105 references)
(1 validation)-
- Key Features
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- 95+ publication references, 1 independent validation
- Frequently used as CUT&RUN IgG negative control and CUT&Tag secondary antibody
- Used in CUT&RUN and CUT&Tag protocols, e.g. Henikoff et al. (2018) PMID 29651053, Kaya-Okur et al. (2019, 2020) PMID 31036827 and PMID 32913232
- 抗原 See all IgG products
- IgG
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抗原表位
- Heavy & Light Chain
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适用
- 兔
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宿主
- 豚鼠
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克隆类型
- 多克隆
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应用范围
- ELISA, Immunohistochemistry (IHC), Western Blotting (WB), Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Cleavage Under Targets and Tagmentation (CUT&Tag)
- 原理
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The Guinea Pig anti-Rabbit IgG antibody ABIN101961 is well suited as a CUT&RUN IgG negative control and as a secondary antibody in CUT&Tag. It is a component of all our CUT&RUN Product Sets.
Find more products for CUT&RUN and CUT&Tag:
- 特异性
- Rabbit IgG (H&L)
- 交叉反应 （详细）
- No reaction was observed against Human, Mouse and Goat Serum Proteins.
- 纯化方法
- Preadsorption: Solid phase absorption
- 过滤
- Sterile filtered
- 免疫原
- whole molecule of rabbit IgG
- 亚型
- IgG
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Goat anti-Rabbit IgG (Heavy & Light Chain) antibody - PreadsorbedELISA, IHC, WB, LF Polyclonal IgG unconjugated
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- 应用备注
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The guinea pig anti-rabbit IgG antibody ABIN101961 is suitable for use in ELISA, immunohistochemistry, and Western Blot, CUT&RUN and CUT&Tag. Specific conditions for each assay should be optimized by the end user. General ABIN101961 dilution recommendations for different applications are as follows:
- ELISA: 1:20,000 - 1:100,000
- WB: 1:2,000 - 1:10,000
- IHC: 1:1,000 - 1:5,000
- CUT&RUN: 1:100
- CUT&Tag: 1:100
- 说明
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ABIN101961 is tested via ELISA to ensure that the titer against the antigen (Rb IgG) is above a certain threshold. We also test to make sure the titer against potentially cross-reactive human IgG, goat IgG, and mouse IgG is below a certain threshold.
In addition, we test ABIN101961 against anti-guinea pig Serum, rabbit IgG, and rabbit serum in an immunoelectrophoresis assay. - 限制
- 仅限研究用
-
- by
- Tom Taghon’s lab, Vakgroep Diagnostische Wetenschappen, Universiteit Gent
- No.
- #104174
- 日期
- 2019.12.18
- 抗原
- rabbit IgG
- Lot Number
- 43586
- Method validated
- Cleavage Under Targets and Tagmentation
- Positive Control
rabbit anti-H3K27me3 monoclonal antibody
- Negative Control
rabbit normal IgG antibody
- Notes
Passed. ABIN101961 successfully increased the number of protein A binding sites in a CUT&Tag protocol on human primary thymocytes and PER-117 cells using a monoclonal rabbit H3K27me3 primary antibody.
- Primary Antibody
- rabbit IgG anti-H3K27me3 antibody (Cell Signaling Technology, 9733T, lot 14)
- Secondary Antibody
- ABIN101961
- Full Protocol
- Profiling of H3K27me3 signals in human primary CD34+ sorted thymocytes and PER-117 cell line (50,000 cells each).
- Carry out the CUT&Tag protocol according to the single-tube bench top protocol for CUT&Tag developed by Steven Henikoff’s lab as outlined on the protocols.io platform.
- Use reagents as suggested in the original protocol. The pA-Tn5 fusion protein used in this validation was a courtesy of Steven Henikoff’s lab and used at 1:1000 dilution for 100 µL per sample.
- Primary antibody binding with
- 1µl/sample monoclonal rabbit IgG anti-H3K27me3 antibody (Cell Signaling Technology, 9733T, lot 14) or
- 1µl/sample rabbit normal IgG (Cell Signaling, 2729S, lot 9).
- Secondary antibody binding with 1µl/sample guinea pig anti rabbit antibody (antibodies-online, ABIN101961, lot 43586).
- Determine library concentration on a Qubit Fluorometer using Qubit dsDNA High Sensitivity Assay Kit (ThermoFisher Scientific, Q32581).
- Determine library size distribution of all samples on an Agilent Fragment Analyzer using High Sensitivity Small DNA Fragment Analysis Kit (Agilent, DNF-477-0500).
- Experimental Notes
ABIN101961 successfully increased the number of protein A binding sites for each bound rabbit anti-H3K27me3 antibody in the human primary thymocytes and PER-117 cell line. This resulted in quantifiable amounts of tagmented genomic fragments after PCR amplification that showed a ladder-like distribution.
- Library concentrations as measured on a Qubit Fluorometer were 0.122ng/µl and 0.538ng/µl for thymocytes and PER-117 cells respectively using a rabbit H3K27me3 primary antibody. Library concentration for both sample was below the instrument’s detections limit using the rabbit normal IgG primary antibody.
- As discussed at the protocols.io, it is expected to observe a ladder-like distribution for H3K27me3 profiling. However, using lower cell numbers could alter the nucleosomal patterns and result in an increase in larger fragments. This is observed by Steven Henikoff’s lab as well. Nevertheless, it does not affect the quality of the sequencing results.
生效 #104174 (Cleavage Under Targets and Tagmentation)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![CUT&Tag library generated from thymocytes using a monoclonal rabbit anti-H3K27me3 primary antibody and ABIN101961 as a secondary antibody.]()
CUT&Tag library generated from thymocytes using a monoclonal rabbit anti-H3K27me3 primary antibody and ABIN101961 as a secondary antibody.
![CUT&Tag library generated from PER-117 cells using a monoclonal rabbit anti-H3K27me3 primary antibody and ABIN101961 as a secondary antibody.]()
CUT&Tag library generated from PER-117 cells using a monoclonal rabbit anti-H3K27me3 primary antibody and ABIN101961 as a secondary antibody.
Full Methods -
- 状态
- Liquid
- 浓度
- 1.21 mg/mL
- 缓冲液
- 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01 % (w/v) NaN3, no stabilizer
- 储存液
- Sodium azide
- 注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 储存条件
- 4 °C,-20 °C
- 有效期
- 12 months
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: "Chem-map profiles drug binding to chromatin in cells." in: Nature biotechnology, (2023) (PubMed).
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- 抗原
- IgG
- Abstract
- IgG 产品
- 物质类
- Antibody
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