Anti-Vimentin antibody [SP20]

• 产品名称

  Anti-Vimentin抗体[SP20]
  参阅全部 Vimentin 一抗

• 描述

  兔单克隆抗体[SP20] to Vimentin

• 宿主

  Rabbit

• 特异性

  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.

• 经测试应用

  适用于: ICC/IF, WB, IHC-P, Flow Cytmore details

• 种属反应性

  与反应: Human
  预测可用于: Rat, Hamster, Cow, Xenopus laevis

• 免疫原

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

• 阳性对照

• 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.

• 常规说明

  This product has switched from a hybridoma to recombinant production method on 4th September 2023.

  This product is a recombinant monoclonal antibody, which offers several advantages including:

  For more information see here.

  
  

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

• - High batch-to-batch consistency and reproducibility

• - Improved sensitivity and specificity

• - Long-term security of supply

• - Animal-free production

• 形式

  Liquid

• 存放说明

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

• 存储溶液

  pH: 7.20
  Preservative: 0.1% Sodium azide
  Constituents: 98.9% PBS, 1% BSA

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  SP20

• 同种型

  IgG

The Abpromise guarantee

Abpromise™承诺保证使用ab16700于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

  hide

• SwissProt: P31000 Rat

• Unigene: 455493 Human

• Unigene: 691131 Human

• Unigene: 2710 Rat

• 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

• 形式

  Vimentin is found in connective tissue and in the cytoskeleton.

• 别名

• HEL113 antibody

• VIM antibody

• VIME_HUMAN antibody

• Vimentin antibody

• CTRCT30 antibody

• Epididymis luminal protein 113 antibody

• FLJ36605 antibody

• 

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

  Immunohistochemical analysis of paraffin-embedded Human melanoma tissue labeling Vimentin with ab16700 at 1/200 dilution. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Western blot - Anti-Vimentin antibody [SP20] (ab16700)

  All lanes : Anti-Vimentin antibody [SP20] (ab16700) at 1/120 dilution
  
  Lane 1 : Wild-type A549 cell lysate
  Lane 2 : Vimentin knockout A549 cell lysate
  Lane 3 : Daudi cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Predicted band size: 53 kDa
  Observed band size: 55 kDawhy is the actual band size different from the predicted?
  
  
  
  

  This image was generated using a previous batch manufactured using the hybridoma production method.

  False colour image of Western blot: Anti-Vimentin antibody [SP20] staining at 1/120 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab16700 was shown to bind specifically to Vimentin. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in VIM knockout cell line ab288984. To generate this image, wild-type and VIM knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

• 

  Western blot - Anti-Vimentin antibody [SP20] (ab16700)

  All lanes : Anti-Vimentin antibody [SP20] (ab16700) at 1/100 dilution
  
  Lane 1 : U20S cell lysate
  Lane 2 : Human 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
  
  
  
  

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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)

  Anti-Vimentin antibody [SP20] (ab16700) at 1/100 dilution + HeLa cell lysate
  
  Predicted band size: 53 kDa
  Observed band size: 53 kDa
  
  
  
  

  This image was generated using a previous batch manufactured using the hybridoma production method.

• 

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

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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).

• 

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

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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)This image is courtesy of an Abreview submitted by Miss Szu-Yu Chen

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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)

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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)

  This image was generated using a previous batch manufactured using the hybridoma production method.

  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.