Vimentin Recombinant Rabbit Monoclonal Antibody [SC60-05]

RMB: 900 特惠 1500 1500

产品规格

50μl

100μl

现在订购，【现货】

点击询价

联系我们

邮箱: sales@huabio.cn

电话: 0571-88062880

查找各地代理商

Catalog# ET1610-39

Vimentin Recombinant Rabbit Monoclonal Antibody [SC60-05]

Application

WB
IHC-P
IHC-Fr
IF-Tissue
IF-Cell
FC
IP

Reactivity

Human
Mouse
Rat

Predicted reactivity

Cynomolgus monkey
Pig

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

Vimentin Recombinant Rabbit Monoclonal Antibody [SC60-05]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within C-terminal human Vimentin.

种属反应性

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

验证应用

WB, IHC-P, IHC-Fr, IF-Tissue, IF-Cell, FC, IP

分子量

Predicted band size: 54 kDa

阳性对照

HeLa cell lysate, HEK-293 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, C6 cell lysate, C2C12 cell lysate, RAW264.7 cell lysate, C6, HeLa, L6, C2C12, human endometrial carcinoma tissue, mouse colon tissue, mouse cerebellum tissue, human breast carcinoma tissue, human colon tissue, human kidney tissue, mouse large intestine tissue.

偶联

unconjugated

克隆号

SC60-05

RRID

AB_3069923

产品特性

形态

Liquid

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

WB
1:20,000-1:50,000

IHC-P
1:5,000-1:10,000

IHC-Fr
1:1,000-1:2,000

IF-Tissue
1:1,000-1:2,000

IF-Cell
1:100-1:500

FC
1:1,000

IP
Use at an assay dependent concentration.

发表文章中的应用

WB	查看 55 篇文献如下
IF	查看 19 篇文献如下
IHC-P	查看 5 篇文献如下
IF-cell	查看 1 篇文献如下
IHC	查看 1 篇文献如下

发表文章中的种属

Human	查看 50 篇文献如下
Mouse	查看 23 篇文献如下
Rat	查看 3 篇文献如下

靶点

功能

Vimentin (57 kDa) is the most ubiquituos intermediate filament protein and the first to be expressed during cell differentiation. All primitive cell types express vimentin but in most non-mesenchymal cells it is replaced by other intermediate filament proteins during differentiation. Vimentin is expressed in a wide variety of mesenchymal cell types: fibroblasts, endothelial cells etc., and in a number of other cell types derived from mesoderm, e.g., mesothelium and ovarian granulosa cells. Vimentin is present in many different neoplasms but is particulary expressed in those originated from mesenchymal cells. Sarcomas e.g., fibrosarcoma, malignt fibrous histiocytoma, angiosarcoma, and leio- and rhabdomyosarcoma, as well as lymphomas, malignant melanoma and schwannoma, are virtually always vimentin positive. Mesoderm derived carcinomas like renal cell carcinoma, adrenal cortical carcinoma and adenocarcinomas from endometrium and ovary usually express vimentin. Also thyroid carcinomas are vimentin positive. Any low differentiated or sarcomatoid carcinoma may express some vimentin. Vimentin is frequently included in the so-called primary panel (together with CD45, cytokeratin, and S-100 protein): Intense staining reaction for vimentin without coexpression of other intermediate filament proteins is strongly suggestive of a mesenchymal tumour or a malignant melanoma. However, in biopsies representing only a sarcomatoid part of renal cell carcinoma a.o. a strong positivity for vimentin without cytokeratin expression may be seen. Tumours like lymphomas and seminomas have the same intermediate filament profile, but the vimentin expression is usually weaker.

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

序列相似性

Belongs to the intermediate filament family.

组织特异性

Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.

翻译后修饰

Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33. Phosphorylated on tyrosine residues by SRMS.; O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.; S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.

亚细胞定位

Cytoplasm.

UNIPROT

SwissProt: P08670 Human

SwissProt: P20152 Mouse

SwissProt: P31000 Rat

别名

CTRCT30 antibody

Epididymis luminal protein 113 antibody

FLJ36605 antibody

HEL113 antibody

VIM antibody

VIME_HUMAN antibody

Vimentin antibody

图片

Application: IHC-Fr

Species: Mouse

Site: Cerebellum

Sample: Frozen section

Antibody concentration: 1/2,000

Antigen retrieval: Not required
Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (ET1610-39) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: HEK-293 cell lysate (10 µg/Lane)
Lane 3: Jurkat cell lysate (10 µg/Lane)
Lane 4: C2C12 cell lysate (10 µg/Lane)
Lane 5: RAW264.7 cell lysate (10 µg/Lane)
Lane 6: C6 cell lysate (10 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 14 seconds; ECL: K1801;

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 (ET1610-39) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution were 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.
Western blot analysis of Vimentin on different lysates with Rabbit anti-Vimentin antibody (ET1610-39) at 1/20,000 dilution.

Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: HEK-293 cell lysate (10 µg/Lane)
Lane 3: Jurkat cell lysate (10 µg/Lane)
Lane 4: NIH/3T3 cell lysate (10 µg/Lane)
Lane 5: C6 cell lysate (10 µg/Lane)
Lane 6: C2C12 cell lysate (10 µg/Lane)
Lane 7: RAW264.7 cell lysate (10 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: Lane 1-5: 3 seconds; Lane 6-7: 14 seconds; ECL: K1801;

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 (ET1610-39) at 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of C6 cells labeling Vimentin with Rabbit anti-Vimentin antibody (ET1610-39) at 1/1,000 dilution and competitor's antibody at 1/200 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-Vimentin antibody (ET1610-39) at 1/1,000 dilution and competitor's antibody at 1/200 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.
Immunocytochemistry analysis of HeLa cells labeling Vimentin with Rabbit anti-Vimentin antibody (ET1610-39) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (ET1610-39) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.
Immunocytochemistry analysis of L6 cells labeling Vimentin with Rabbit anti-Vimentin antibody (ET1610-39) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Vimentin antibody (ET1610-39) at 1/50 dilution in 2% negative goat serum 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.
☑ Knockout (KO)

All lanes: Western blot analysis of Vimentin with anti-Vimentin antibody (ET1610-39) at 1/5,000 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2: Vimentin knockout Hela whole cell lysate (10 µg).

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-39, 1/5,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1/300,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 mouse cerebellum tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 rat colon tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 rat cerebellum tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 breast carcinoma tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/10,000 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 (ET1610-39) at 1/10,000 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 endometrial carcinoma tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 colon tissue with Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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 Rabbit anti-Vimentin antibody (ET1610-39) at 1/5,000 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 (ET1610-39) at 1/5,000 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.
☑ Relative expression (RE)

Flow cytometric analysis of HeLa (positive, red) and MCF7 (negative, green) cells labeling Vimentin.

Cells were fixed by 4% formaldehyde and then permeabilized with ice-cold 90% methanol. Then stained with the primary antibody (ET1610-39) at 1/1,000 dilution, compared with Rabbit IgG Isotype Control (HeLa black, MCF7 light 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℃.
Vimentin was immunoprecipitated in 0.2mg HeLa cell lysate with ET1610-39 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1610-39 at 1/10,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: Rabbit IgG instead of ET1610-39 in HeLa cell lysate
Lane 3: ET1610-39 IP in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 5 seconds
Application: IF-tissue

Species: Mouse

Site: Kindey

Sample: Paraffin-embedded section

Antibody concentration: 1/1,000
Application: IF-tissue

Species: Mouse

Site: Cerebral cortex

Sample: Paraffin-embedded section

Antibody concentration: 1/1,000