PRODUCT CODE: EM0401

Vimentin Mouse Monoclonal Antibody [D4-B11] (EM0401)

  • IVD–IHC

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Vimentin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM0401, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A549 cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 2: NIH/3T3 cell lysate<br />
Lane 4: Mouse embryonic stem cell lysate
  • Western blot analysis of Vimentin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM0401, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A549 cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 2: NIH/3T3 cell lysate<br />
Lane 4: Mouse embryonic stem cell lysate
  • Western blot analysis of Vimentin on normal (Hela Control) and Vimentin knockolut Hela lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM0401, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Vimentin in Hela cells (red). 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 (EM0401, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 using anti-Vimentin antibody. 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 (EM0401, 1/800) 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 using anti-Vimentin antibody. 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 (EM0401, 1/800) 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.
Western blot analysis of Vimentin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM0401, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A549 cell lysate
Lane 2: Hela cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 4: Mouse embryonic stem cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

Vimentin Mouse Monoclonal Antibody [D4-B11] (EM0401)

Immunogen

Peptide

Host

Mouse

Positive Control

A549, Hela, NIH/3T3, mouse embryonic stem cell human lung?tissue, human colon carcinoma tissue, human stomach carcinoma tissue, mouse lung tissue

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

D4-B11

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

54 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:2,000-1:5,000

  • ICC

  • 1:200

  • IHC-P

  • 1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Vimentin

SYNONYMS

CTRCT30 antibody; Epididymis luminal protein 113 antibody; FLJ36605 antibody; HEL113 antibody; VIM antibody; VIME_HUMAN antibody; Vimentin antibody

SEQUENCE SIMILARITIES

Belongs to the intermediate filament family.

TISSUE SPECIFICITY

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.

POST-TRANSLATIONAL MODIFICATION

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.

SUBCELLULAR LOCATION

Cytoplasm

FUNCTION

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.