Synthetic peptide within Human Vimentin aa 1-50 / 466.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IHC-P
分子量
Predicted band size: 54 kDa
阳性对照
HeLa cell lysate, C2C12 cell lysate, L6 cell lysate, HeLa, human appendix tissue, human kidney tissue, human liver tissue.
偶联
unconjugated
克隆号
A6-C1-R
产品特性
形态
Liquid
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG1
纯化方式
Protein A affinity purified.
应用稀释度
WB
1:2,000
IF-Cell
1:100
IHC-P
1:4,000
靶点
功能
Vimentin is a type III intermediate filament (IF) protein that is expressed in mesenchymal cells. Vimentin plays a significant role in supporting and anchoring the position of the organelles in the cytosol. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally. In essence, vimentin is responsible for maintaining cell shape, integrity of the cytoplasm, and stabilizing cytoskeletal interactions. Vimentin has been shown to eliminate toxic proteins in JUNQ and IPOD inclusion bodies in asymmetric division of asymmetric division of mammalian cell lines. It has been used as a sarcoma tumor marker to identify mesenchyme. Methylation of the vimentin gene has been established as a biomarker of colon cancer and this is being utilized in the development of fecal tests for colon cancer. High levels of DNA methylation in the promotor region have also been associated with markedly decreased survival in hormone positive breast cancers.
背景文献
1. Ridge KM et al. Roles of vimentin in health and disease. Genes Dev. 2022 Apr
2. Kuburich NA et al. Vimentin and cytokeratin: Good alone, bad together. Semin Cancer Biol. 2022 Nov
Western blot analysis of Vimentin on different lysates with Mouse anti-Vimentin antibody (HA610126) at 1/2,000 dilution.
Lane 1: HeLa cell lysate Lane 2: C2C12 cell lysate Lane 3: L6 cell lysate Lane 4: Daudi cell lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 54 kDa Observed band size: 54 kDa
Exposure time: 1 minute 21 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 (HA610126) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells labeling Vimentin with Mouse anti-Vimentin antibody (HA610126) 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 Mouse anti-Vimentin antibody (HA610126) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
Immunohistochemical analysis of paraffin-embedded human appendix tissue with Mouse anti-Vimentin antibody (HA610126) at 1/4,000 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610126) at 1/4,000 dilution for 1 hour 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 kidney tissue with Mouse anti-Vimentin antibody (HA610126) at 1/4,000 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610126) at 1/4,000 dilution for 1 hour 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 Mouse anti-Vimentin antibody (HA610126) at 1/4,000 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610126) at 1/4,000 dilution for 1 hour 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.
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