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    vimentin-antibody-sp20-ab16700.pdf

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Validated using a knockout cell lineRabMAb

Anti-Vimentin antibody [SP20] (ab16700)

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  • SDS
Reviews (7) Submit a question References (76)

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Western blot - Anti-Vimentin antibody [SP20] (ab16700)
  • Western blot - Anti-Vimentin antibody [SP20] (ab16700)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
  • Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)
  • Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)

Key features and details

  • Rabbit monoclonal [SP20] to Vimentin
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

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Human VIM (Vimentin) knockout HeLa cell line (ab255446)

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Overview

  • Product name

    Anti-Vimentin antibody [SP20]
    See all Vimentin primary antibodies
  • Description

    Rabbit monoclonal [SP20] to Vimentin
  • Host species

    Rabbit
  • Specificity

    We have data to show that ab16700 is not suitable for work on mouse tissue. For researchers working on mouse we recommend using ab92547. If you would like further information on this, please do not hesitate to contact our technical support team.
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat, Hamster, Cow, Xenopus laevis
  • Immunogen

    Recombinant full length protein corresponding to Human Vimentin aa 1 to the C-terminus.

  • Positive control

    • WB: U-2 OS, Hu tonsil and HeLa cell lysates. Flow Cyt: HeLa cells. ICC/IF: HAP1-VIM cells, human limbal epithelial cells. IHC-P: Human breast cancer and melanoma tissue. IHC-Fr: Colorectal cancer tissue.
  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.50
    Preservative: 0.1% Sodium azide
    Constituents: Tissue culture supernatant, Tris buffered saline, 1% BSA
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    SP20
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neural Stem Cell marker
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class III
    • Vimentin
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Lineage Markers
    • Endoderm
    • Stem Cells
    • Neural Stem Cells
    • Intracellular
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Neurology process
    • Neuroregeneration
    • Neuroregeneration
    • Developmental Biology
    • Lineage specification
    • Ectoderm
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Vimentin antibody [SP20], prediluted (ab27608)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human VIM (Vimentin) knockout HeLa cell line (ab255446)
  • KO cell lysates

    • Human VIM (Vimentin) knockout HeLa cell lysate (ab263775)
  • KO cell pellets

    • Human VIM (Vimentin) knockout HeLa cell pellet (ab278921)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
  • Related Products

    • Human Vimentin Profiling ELISA Kit (ab173190)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab16700 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF (3)
1/1000.
WB (1)
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-P
1/200.

Antigen Retrieval: None

Flow Cyt
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
ICC/IF
1/1000.
WB
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-P
1/200.

Antigen Retrieval: None

Flow Cyt
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
    Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
  • 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.
  • Involvement in disease

    Cataract 30
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Domain

    The central alpha-helical coiled-coil rod region mediates elementary homodimerization.
    The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
  • Post-translational
    modifications

    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.
    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.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P08670 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 280955 Cow
    • Entrez Gene: 101824289 Hamster
    • Entrez Gene: 7431 Human
    • Entrez Gene: 81818 Rat
    • Omim: 193060 Human
    • SwissProt: P48616 Cow
    • SwissProt: P02544 Hamster
    • SwissProt: P08670 Human
    • SwissProt: P31000 Rat
    • Unigene: 455493 Human
    • Unigene: 691131 Human
    • Unigene: 2710 Rat
    see all
  • Form

    Vimentin is found in connective tissue and in the cytoskeleton.
  • Alternative names

    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Images

  • Western blot - Anti-Vimentin antibody [SP20] (ab16700)
    Western blot - Anti-Vimentin antibody [SP20] (ab16700)
    All lanes : Anti-Vimentin antibody [SP20] (ab16700)

    Lane 1 : U20S cell lysate
    Lane 2 : Hu tonsil cell lysate
    Lane 3 : Wild-type HeLa cell lysate
    Lane 4 : VIM knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 53 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab16700 observed at 53 kDa. Red - loading control, ab8245 observed at 37 kDa.

     ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Vimentin antibody [SP20] (ab16700)
    Western blot - Anti-Vimentin antibody [SP20] (ab16700)
    Anti-Vimentin antibody [SP20] (ab16700) at 1/100 dilution + HeLa cell lysate

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)

    ab16700 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab16700 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)

    Immunohistochemical analysis of Human melanona tissue labelling Vimentin with ab16700.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20] (ab16700)

    IHC image of ab16700 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16700, (Neat supernatant) for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)This image is courtesy of an anonymous Abreview

    ab16700 staining Vimentin in human corneal limbal epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.3% Triton X-100 for 5 minutes. Samples were incubated with primary antibody (1/200 in PBS + 10% normal goat serum) for 18 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)
    Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20] (ab16700)This image is courtesy of an Abreview submitted by Miss Szu-Yu Chen

    ab16700 at 1/200 staining Human Limbal Epithelial Cells by ICC/IF. The cells were incubated with the antibody for 1 hour and then a FITC conjugated goat antibody was used as the secondary. The image shows vimentin staining in green and hoechst staining in blue. The upper cells in the image (vimentin negative) are epithelium cells. the vimentin positive cells are stroma cells.

    See Abreview

  • Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)
    Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)

    Flow cytometric analysis of rabbit anti-Vimentin (SP20) antibody ab16700 (1/100) in HeLa cells (green) compared to negative control of rabbit IgG (blue).

  • Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)
    Flow Cytometry - Anti-Vimentin antibody [SP20] (ab16700)

    Overlay histogram showing HeLa cells stained with ab16700 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16700, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (76)

Publishing research using ab16700? Please let us know so that we can cite the reference in this datasheet.

ab16700 has been referenced in 76 publications.

  • Chaney HL  et al. Conceptus-induced, interferon tau-dependent gene expression in bovine endometrial epithelial and stromal cells†. Biol Reprod 104:669-683 (2021). PubMed: 33330929
  • Lombardi R  et al. HSP90 identified by a proteomic approach as druggable target to reverse platinum resistance in ovarian cancer. Mol Oncol 15:1005-1023 (2021). PubMed: 33331136
  • Xiong G  et al. Circular RNA_0074027 participates in cell proliferation, apoptosis and metastasis of colorectal cancer cells through regulation of miR-525-3p. Mol Med Rep 23:N/A (2021). PubMed: 33760126
  • Liu Y  et al. Role of microRNA-25 in high glucose cultured Müller glia. Int J Ophthalmol 14:643-648 (2021). PubMed: 34012877
  • Wang G  et al. The pan-cancer landscape of crosstalk between epithelial-mesenchymal transition and immune evasion relevant to prognosis and immunotherapy response. NPJ Precis Oncol 5:56 (2021). PubMed: 34158591
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review

Filter by Application

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1-7 of 7 Abreviews

Immunocytochemistry/ Immunofluorescence abreview for Anti-Vimentin antibody [SP20]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDA-MB-231)
Permeabilization
Yes - 0.5% Triton X for 15 min
Specification
MDA-MB-231
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Jorge Guerrero

Verified customer

Submitted Jun 19 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Vimentin antibody [SP20]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Corneal limbus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA (ph 9.0)
Permeabilization
Yes - PBT: 0.1% Triton X-100 in PBS
Specification
Corneal limbus
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Noelia Andollo

Verified customer

Submitted Oct 08 2018

Immunohistochemistry (Frozen sections) abreview for Anti-Vimentin antibody [SP20]

Poor
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Bryan Niedenberger

Verified customer

Submitted Jul 12 2016

Western blot abreview for Anti-Vimentin antibody [SP20]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Dog Cell lysate - whole cell (Osteosarcoma cell lines)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Specification
Osteosarcoma cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Holly Pondenis

Verified customer

Submitted Nov 21 2015

Immunohistochemistry (Frozen sections) abreview for Anti-Vimentin antibody [SP20]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Pancreas)
Permeabilization
No
Specification
Pancreas
Blocking step
CAS block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 09 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Vimentin [SP20] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Corneal limbal epithelial cells)
Specification
Corneal limbal epithelial cells
Permeabilization
Yes - Triton 0,3% - 5 minutes
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 13 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Vimentin antibody [SP20]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Limbal Stromal cells)
Specification
Human Limbal Stromal cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MISS. Ashley Quintana

Verified customer

Submitted Nov 20 2007

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