Invitrogen

HP1 gamma Monoclonal Antibody (2MOD-1G6)

高级验证

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

查看其他19个HP1 gamma抗体

说明书

Protocols

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说明书

Protocols

技术支持

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HP1 gamma Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of HP1-gamma (green) in HeLa cells. The cells were fixed with formalin for 15 minutes, permeabilized with 1X Permeablilization buffer (Product # 8408400) for 15 minutes, and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were stained with HP1-gamma monoclonal antibody (Product # MA3-054) at a dilution of 1:250 in blocking buffer for one hour at room temperature, washed with 1X TBS Tween 20 Buffer (Product # 28360), followed by incubation with DyLight 488 goat anti-mouse IgG secondary antibody (green, Product # A28175) at a dilution of 1:2000 and Hoechst 33342 dye (blue, Product # 62249) at a dilution of 1:5000 for 30 minutes at room temperature. Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

HP1 gamma Antibody in Western Blot (WB) — Western blot analysis of HP1-gamma was performed by loading 20 µg of NIH 3T3 (lane 1), MEF (lane 2), and NRK (lane 3) cell lysates in reducing sample buffer (Product # 39000) and Page Ruler Plus Protein Ladder (Product # 26619) onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4201BX10). Proteins were transferred to nitrocellulose membrane (Product # 88018) with Transfer Buffer (Product # 84731) using the G2 Fast Blotter (Product # 62288). Membrane was blocked in StartingBlock T20 (Product # 37543) for one hour at room temperature. HP1-gamma was detected at approximately 22 kDa using a HP1-gamma monoclonal antibody (Product # MA3-054) at a dilution of 1:2000 in StartingBlock T20 overnight at at 4°C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236).

HP1 gamma Antibody in ChIP assay (ChIP) — Chromatin immunoprecipitation analysis of HIV1 HP1-gamma was performed using cross-linked chromatin from Human 5A8 J-lat T lymphocytes culture latently infected with HIV1 and treated with 10 µg/mL PHA (phytohemaglutin) for 0, 4, and 8 hours. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of HP1-gamma monoclonal antibody (Product # MA3-054). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers shown to amplify exon-1 of the Egr1 gene or HIV gag gene. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. Schematic representations of the Egr1 and HIV1 gag locus are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by the primers are represented by black bars. Data courtesy of the Innovators Program.

HP1 gamma Antibody in ELISA (ELISA) — ELISA analysis of HP1-gamma was performed by coating wells of a 96-well plate with 100 µL per well of recombinant HP1-gamma protein or BSA (Product # 23209) diluted in carbonate/bicarbonate buffer (Product # 28382), at a concentration of 1 µg/mL. Wells of the plate were washed, blocked with Superblock blocking buffer (Product # 37536), and incubated with 100 µL per well of a mouse anti-HP1-gamma monoclonal antibody (Product # MA3-054) starting at a concentration of 1 µg/mL and serially diluting 2-fold to a concentration of 1 ng/mL for 1 hour at 37°C. The plate was washed, then incubated with 100 µL per well of goat anti-mouse IgG SuperclonalTM secondary antibody (Product # A28174) at a dilution of 1:10,000 for 1 hour at 37°C followed by washing. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550 nm.

HP1 gamma Antibody — The specificity of anti-CBX3/HP1 gamma mouse monoclonal antibody (Product # MA3-054) was demonstrated by CRISPR targeted CBX3/HP1 gamma knockout in HeLa cells. Western blot analysis of whole cell lysates using this antibody showed no detection of CBX3/HP1 gamma protein expression in knockout cells (lane 1) compared to the CBX3/HP1 gamma protein detected at ~20kDa in wildtype HeLa cells (lane 2). {KO}

产品信息

MA3-054

应用	建议稀释比	已发表文章
免疫印迹 (WB)	1:2,000	-
免疫细胞化学 (ICC/IF)	1:250	-
酶联免疫吸附实验 (ELISA)	1:500	-
染色质免疫沉淀实验 (ChIP)	1:100	-

产品规格
种属反应	Human, Mouse, Rat
宿主/亚型	Mouse / IgG1
分类	Monoclonal
类型	Antibody
克隆号	2MOD-1G6
抗原	Recombinant HP1-gamma mouse
偶联物	Unconjugated
形式	Liquid
浓度	Conc. Not Determined
保存液	ascites
内含物	0.05% sodium azide
保存条件	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2607959

产品详细信息

MA3-054 detects HP1-gamma in human, mouse and rat samples.

MA3-054 has been succesfully used in western blot, ELISA, immunofluorescence and ChIP applications.

Immunogen is a HP1-gamma recombinant mouse

靶标信息

At the nuclear envelope, the nuclear lamina and heterochromatin are adjacent to the inner nuclear membrane. The protein encoded by this gene binds DNA and is a component of heterochromatin. This protein also can bind lamin B receptor, an integral membrane protein found in the inner nuclear membrane. The dual binding functions of the encoded protein may explain the association of heterochromatin with the inner nuclear membrane.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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生物信息学

蛋白别名: chromobox homolog 3; chromobox homolog 3 (HP1 gamma homolog, Drosophila); Chromobox protein homolog 3; HECH; heterochromatin protein 1 gamma; Heterochromatin protein 1 homolog gamma; heterochromatin protein HP1 gamma; heterochromatin-like protein 1; HGNC:1553; HP1 gamma; HP1 gamma homolog; HPy; Modifier 2 protein; OTTHUMP00000202463

基因别名: CBX3; HECH; HP1-GAMMA; HP1g; HP1Hs-gamma; M32

UniProt ID: (Human) Q13185

Entrez Gene ID: (Human) 11335, (Mouse) 12417, (Rat) 297093

功能

chromatin remodeling transcription, DNA-templated covalent chromatin modification negative regulation of transcription, DNA-templated rhythmic process cellular response to dexamethasone stimulus regulation of transcription, DNA-templated chromatin modification

参与通路

protein binding enzyme binding protein domain specific binding identical protein binding histone methyltransferase binding transcription regulatory region sequence-specific DNA binding molecular_function

HP1 gamma Monoclonal Antibody (2MOD-1G6)

产品信息

MA3-054

免疫印迹 (WB)

免疫细胞化学 (ICC/IF)

酶联免疫吸附实验 (ELISA)

染色质免疫沉淀实验 (ChIP)

种属反应

宿主/亚型

分类

类型

克隆号

抗原

偶联物

形式

浓度

保存液

内含物

保存条件

RRID

产品详细信息

靶标信息

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HP1 gamma Monoclonal Antibody (2MOD-1G6)

产品信息

MA3-054

免疫印迹 (WB)

免疫细胞化学 (ICC/IF)

酶联免疫吸附实验 (ELISA)

染色质免疫沉淀 实验 (ChIP)

种属反应

宿主/亚型

分类

类型

克隆号

抗原

偶联物

形式

浓度

保存液

内含物

保存条件

RRID

产品详细信息

靶标信息

引用文献 (0)

生物信息学

性能保障

随时为您服务

Brands

Shopping Tools

eSolutions

Support

我们的公司

Your items have has been added!

染色质免疫沉淀实验 (ChIP)