Phospho-Hsp27 (Ser78) 抗体 (HY-P80823)
HSPB1; HSP27; HSP28; Heat shock protein beta-1; HspB1; 28 kDa heat shock protein; Estrogen-regulated 24 kDa protein; Heat shock 27 kDa protein; HSP 27; Stress-responsive protein 27; SRP27
反应种属：Human
抗原名称：Phospho-Hsp27 (Ser78)
应用：WB, IF/ICC, IHC-P
标记：Unconjugated
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经验证应用
| 项目 | 详情 |
|---|---|
| WB | Western blot analysis was performed on protein extracts (25 μg) from HeLa (lane 2, Negative), HeLa (lane 3, Calyculin A ( HY-18983 ), 10 nM, 15 mins), HeLa (lane 4, Calyculin A ( HY-18983 ), 25 nM, 15 mins), HeLa (lane 5, Calyculin A ( HY-18983 ), 50 nM, 15 mins), and THP-1 (lane 6) using Phospho-Hsp27 (Ser78) antibody. Proteins were transferred onto a 0.22 μm PVDF membrane using a wet transfer system at a voltage of 350 mA for 90 mins. The membrane was then blocked with 5% BSA in TBST ( HY-K1025 ) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) ( HY-P80954A ) (1:5000) were diluted in 5% BSA in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit ( HY-K1005 ). |
| IHC-P | Immunohistochemical analysis of paraffin-embedded human Colon cancer tissue using Phospho-Hsp27 (Ser78) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80823, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium. Immunohistochemical analysis of paraffin-embedded human Breast Cancer tissue using Phospho-Hsp27 (Ser78) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80823, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium. Immunohistochemical analysis of paraffin-embedded human Ovarian Cancer tissue using Phospho-Hsp27 (Ser78) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80823, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium. Immunohistochemical analysis of paraffin-embedded human Bladder cancer tissue using Phospho-Hsp27 (Ser78) antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80823, 1:100 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium. Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human renal cancer tissue using Phospho-Hsp27 (Ser78) Antibody (HY-P80823, 1/100). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
| ICC | Immunocytochemistry analysis of HeLa cells labeling Phospho-Hsp27 (Ser78) with Phospho-Hsp27 (Ser78) Antibody ( HY-P80823 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Phospho-Hsp27 (Ser78) Antibody ( HY-P80823 ) at 1/50 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of MCF-7 cells labeling Phospho-Hsp27 (Ser78) with Phospho-Hsp27 (Ser78) Antibody ( HY-P80823 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Phospho-Hsp27 (Ser78) Antibody ( HY-P80823 ) at 1/50 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). |
推荐稀释比例
- WB1:500-1:1000
- IHC-P1:50-1:100
- ICC/IF1:50-1:200
详细信息
免疫原Synthetic phosphopeptide corresponding to residues surrounding Ser78 of Human HSP27.The exact sequence is proprietary to MCE.
宿主/亚型Rabbit / IgG
抗体类型Polyclonal
全称Phospho-Hsp27 (Ser78) 抗体
理论分子量23 kDa
实际/观测分子量27 kDa 请注意：因蛋白存在修饰或聚体等情况，以实测为准，预测仅为参考。
产品形式溶液
偶联物Non-conjugated
储存缓冲液Supplied in 1*PBS (pH 7.3), 50% glycerol and 0.5% BSA. Preservative: 0.02% sodium azide.
储存条件保存条件 & 期限: Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
运输条件: Shipping with blue ice.
纯化方式affinity purified
Gene ID3315 [ NCBI ]
RRIDAB_3102211
UniProt IDP04792
背景信息
功能： Hsp27 是一种小热休克蛋白，作为分子伴侣发挥功能，可能将变性蛋白维持在可折叠状态。Hsp27 在应激抵抗和肌动蛋白组装中发挥作用。通过其分子伴侣活性，Hsp27 可调控多种生物学过程，包括神经丝蛋白的磷酸化和轴突运输 [1] [2] [3] [4] 。 亚细胞定位： 细胞质；细胞核；细胞质、细胞骨架、纺锤体 表达水平： 组织特异性：在所有检测的组织中均检测到该物质：骨骼肌、心脏、主动脉、大肠、小肠、胃、食管、膀胱、肾上腺、甲状腺、胰腺、睾丸、脂肪组织、肾脏、肝脏、脾脏、大脑皮层、血清和脑脊液。心脏以及由横纹肌和平滑肌组成的组织中含量最高。 诱导：在热休克等环境胁迫下表达上调 (PubMed:8325890) 。雌激素刺激可上调表达 (PubMed:2743305) ； (微生物感染) 在肠道病毒71型 (EV71) 感染后表达上调 (蛋白水平) 。 亚基： 同源寡聚体 (PubMed:10383393)。同源二聚体；激活后转化为单体 (PubMed:20178975)。异源寡聚体；与 HSPB6 相互作用 (PubMed:23948568)。与 α- 和 β-微管蛋白结合 (PubMed:10777697)。与 TGFB1I1 相互作用 (基于相似性)。与 CRYAB 相互作用 (PubMed:1560006)。与 HSPB8 相互作用 (PubMed:11342557)。与 HSPBAP1 相互作用 (PubMed:10751411)。