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Rabbit抗原名称
Phospho-Histone H3 (Ser28)分子别名
H3S28P免疫原
Synthetic Peptide细胞定位
NucleusAccession
P68431克隆号
S-1025-45抗体类型
Recombinant mAb抗体同种型
IgG翻译后修饰类型
磷酸化反应种属 ?
Hu, Ms, Rt纯化方式
Protein A浓度
0.5 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
储存条件
12 months from date of receipt / reconstitution, -20 °C as supplied
应用
ChIP
IHC-P ?
WB
IP
稀释度
应用 稀释度 推荐种属 Dot Blot 1:1000 WB 1:1000 Hu IP 1:50 Hu IHC-P 1:250 Hu, Ms, Rt ChIP 1:20-1:50 Hu
H3S28P is an epigenetic modification to the DNA packaging protein histone H3. It is a mark that indicates the phosphorylation the 28th serine residue of the histone H3 protein. Early mitosis phosphorylation patterns for H3S10 and H3S28 are quite similar, starting at the initiation of chromosomal condensation during prophase. At gene promoters, H3S28 phosphorylation is expected to remove Polycomb restrictive complexes from chromatin, causing demethylation and acetylation of the nearby K27 residue, so activating transcription. This transition is mediated by the phosphorylation of H3S28 on a site next to H3K27. It was discovered that phosphorylating H3S28 in the promoters of genes like c-fos and -globin controlled their activation.
免疫印迹
WB result of Phospho-Histone H3 (Ser28) Recombinant Rabbit mAb
Primary antibody: Phospho-Histone H3 (Ser28) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 100 ng/ml Colcemid for 16 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 14 kDa
免疫沉淀
Phospho-Histone H3 (Ser28) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Phospho-Histone H3 (Ser28) in 0.4 mg HeLa treated with 100 ng/ml Colcemid for 16 hours whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-Histone H3 (Ser28) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa treated with 100 ng/ml Colcemid for 16 hours whole cell lysate 20 µg (Input)
Lane 2: Phospho-Histone H3 (Ser28) Rabbit mAb IP in HeLa treated with 100 ng/ml Colcemid for 16 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa treated with 100 ng/ml Colcemid for 16 hours whole cell lysate
Predicted MW: 15 kDa
Observed MW: 14 kDa
斑点杂交
Dot blot result of Phospho-Histone H3 (Ser28) Recombinant Rabbit mAb
Lane 1: Phospho-Histone H3 (Ser28) peptide
Lane 2: Histone H3 unmodified peptide
Primary antibody: Phospho-Histone H3 (Ser28) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
免疫组化
IHC shows positive staining in paraffin-embedded human colon. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse colon. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat colon. Anti- Phospho-Histone H3 (Ser28) antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
免疫细胞化学
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa + Nocodazole (100ng/ml,18h) cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used Phospho-Histone H3 (Ser28) Recombinant Rabbit mAb (S-1025-45) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, GAPDH, AFM, MYOD1,
SAT-α and SAT-2 in Phospho-Histone H3 (Ser28)
Recombinant Rabbit mAb (S-1025-45)-immunoprecipitated
sample.
FAQs
我们一般不推荐客户回收利用抗体。 因为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,我们对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
我们推荐客户使用TPST+5%脱脂奶粉来稀释一抗,进行封闭。 虽然BSA被推荐为WB检测磷酸化蛋白的常用封闭剂,但是脱脂奶粉获取更加方便,覆盖更广泛的非特异性结合位点,在一抗性能优越的前提下,使用脱脂奶粉封闭性价比更高







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