Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb (A22146)at 1:5000 dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Negative control (NC): H3 prokaryotic proteinExposure time: 45s.
Confocal imaging of PC-12 cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
Confocal imaging of NIH/3T3 cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
Confocal imaging of HeLa cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
Dot-blot analysis of all sorts of peptides using TriMethyl-Histone H3-K4 antibody (A22146) at 1:50000 dilution.
Immunohistochemistry analysis of paraffin-embedded Human colon tissue using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Mouse esophagus tissue using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat skin tissue using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Human cervix tissue using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K4 antibody (A22146) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from 293T using 5 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K4 in the representative genomic region surrounding RPL30 gene.
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from 293T using 5 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina (RK20265) from 10⁵ K-562 cells with 1 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146), followed by incubation with Goat Anti-Rabbit IgG(H+L)(AS070). The CUT&Tag results denote the enrichment pattern of TriMethyl-Histone H3-K4 around RPL30 gene.
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina (RK20265) from 10⁵ K-562 cells with 1 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146), followed by incubation with Goat Anti-Rabbit IgG (H+L)(AS070). The CUT&Tag results denote the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).
| WB (Western Blot) | Human, Mouse, Other |
|---|---|
| IF (Immunofluorescence) | Human |
| ChIP (Chromatin Immunoprecipitation) | Monkey, Arabidopsis |
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Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.
首先，一般抗体不推荐客户回收利用，抗体使用之后缓冲体系已经发生改变，不同客户在回收抗体的保存条件上也会有差异，所以抗体回收使用效果无法保证。另外，ABclonal公司也做过一批抗体回收验证测试，测试结果显示不同抗体可回收次数不同，一般效价越高的抗体，可重复使用的次数越多，客户可根据实验情况来确定。
注：我们将孵育完毕后剩余的抗体回收到离心管中置于4℃保存，效价高的抗体可至少保存1周，至少重复利用3次。
武汉爱博泰克生物（ABclonal）科技有限公司是国产品牌，她成立于2011年，公司依托ABclonal美国波士顿抗体与蛋白研发中心、中国光谷生物城(武汉)抗体生产基地以及上海张江分子酶研发中心，凝聚了十余位来自哈佛大学、麻省理工、复旦大学、上海交大、中科院生化细胞所和武汉大学的全球知名分子和免疫学方面博士，组成我们的科学家团队，通过聚焦抗体与酶核心技术，致力于打破国际技术的垄断，将公司打造成为科研工具和诊断原料的国内领导品牌，乃至弯道超越国际巨头。 我们拥有包括兔多克隆抗体、小鼠单克隆抗体、兔单克隆抗体的生产研发平台，同时也有包括WB,IHC,IF,IP,CHIP在内的检测平台，我们对每一支自产的抗体进行了严格的检测。当然，我们部分直销地区也可以帮客户代购进口品牌的产品。同时也有抗体定制服务。ABclonal抗体优势：1，严自检，保质量；2产品多，指标全；3，价格低，货期短。注：ABclonal抗体价格体系详情见附件
ABclonal抗体成分在平时工作当中，常会有客户咨询我们的抗体用的什么buffer进行保存，一般来说，我们的buffer的成分是：PBS含0.03%的proclin300、0.05%牛血清白蛋白、50%甘油；也有一些是PBS含0.03%的proclin300，50%甘油。防腐剂 Proclin 300活性成分主要是2-甲基-4-异噻唑啉-3-酮（MCI）和5-氯-2-甲基-4-异噻唑啉-3-酮（CMCI）。ProClin生物灭活剂能够迅速穿透细胞膜，抑制对细胞呼吸至关重要的特定酶，因此一接触微生物有机体就会立即抑制细胞活性。ProClin的多个特定毒性位点可以防止微生物产生高水平的耐药性。
