Cytokeratin 18 Recombinant Rabbit Monoclonal Antibody [SZ80-07] - BSA and Azide free
Catalog# HA750069
Cytokeratin 18 Recombinant Rabbit Monoclonal Antibody [SZ80-07] - BSA and Azide free
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WB
-
IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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ET1603-8
含抗保成分
-
unconjugated
概述
产品名称
Cytokeratin 18 Recombinant Rabbit Monoclonal Antibody [SZ80-07] - BSA and Azide free
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Recombinant protein within mouse Cytokeratin 18 aa 200-423.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P, FC
分子量
Predicted band size: 48 kDa
阳性对照
A431 cell lysate, HeLa cell lysate, HT-29 cell lysate, mouse kidney, rat skin tissue, Hela, A431, HepG2, human colon carcinoma tissue, human breast carcinoma tissue, human colon tissue,human liver tissue, human stomach tissue, human stomach carcinoma tissue, MCF-7.
偶联
unconjugated
克隆号
SZ80-07
产品特性
形态
Liquid
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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WB
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1:5,000-1:20,000
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IF-Cell
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1:100-1:500
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IF-Tissue
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1:100-1:500
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
靶点
功能
Cytokeratins comprise a diverse group of intermediate filament proteins (IFPs) that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. Cytokeratins have been found to be useful markers of tissue differentiation which is directly applicable to the characterization of malignant tumors. For example, Cytokeratins 10 and 13 are expressed highly in a subset of squamous cell carcinomas while Cytokeratin 18 is expressed in a majority of adenocarcinomas and basal cell carcinomas. Cytokeratin 18 contains two major phosphorylation sites on Ser 33 and Ser 52. Phosphorylation of Ser 18 is essential for the association of Cytokeratin 18 with 14-3-3 proteins and is involved in keratin organization and distribution.
背景文献
1. Kuramoto G et al. Preventive effect of oral mucosal epithelial cell sheets on intrauterine adhesions. Hum Reprod 30:406-16 (2015).
2. Huang YS et al. RGD surface functionalization of the hydrophilic acrylic intraocular lens material to control posterior capsular opacification. PLoS One 9:e114973 (2014).
序列相似性
Belongs to the intermediate filament family.
组织特异性
Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
翻译后修饰
Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.; Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.; O-GlcNAcylation increases solubility, and decreases stability by inducing proteasomal degradation.
亚细胞定位
Cytoplasm, Nucleus.
别名
Cell proliferation inducing gene 46 protein antibody
Cell proliferation inducing protein 46 antibody
Cell proliferation-inducing gene 46 protein antibody
CK 18 antibody
CK-18 antibody
CK18 antibody
CYK 18 antibody
CYK18 antibody
Cytokeratin 18 antibody
Cytokeratin endo B antibody
展开图片
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Western blot analysis of Cytokeratin 18 on different lysates with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/5,000 dilution.
Lane 1: A431 cell lysate
Lane 2: HeLa cell lysate
Lane 3: HT-29 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 48 kDa
Observed band size: 48 kDa
Exposure time: 24 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750069) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of Cytokeratin 18 on different lysates with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/20,000 dilution.
Lane 1: HeLa-si NT cell lysate
Lane 2: HeLa-si Cytokeratin 18 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 48 kDa
Observed band size: 48 kDa
Exposure time: 5 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750069) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunofluorescence analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 18 with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750069, red) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
Immunocytochemistry analysis of A431 cells labeling Cytokeratin 18 with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/100 dilution.
Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750069, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
Immunocytochemistry analysis of HepG2 cells labeling Cytokeratin 18 with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/100 dilution.
Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750069, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/50) for 30 minutes at room temperature. 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 breast carcinoma tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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 colon tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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 stomach tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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 colon carcinoma with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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 stomach carcinoma tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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 tissue with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750069, 1/200) for 30 minutes at room temperature. 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. -
Immunocytochemistry analysis of HeLa cells labeling Cytokeratin 18 with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 18 antibody (HA750069) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HeLa cells labeling Cytokeratin 18.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750069, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Application: IF-Tissue
Species: Human
Site: liver
Sample: Paraffin-embedded section
Antibody concentration: 1/200
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