Anti-Vimentin 抗体 [SP20] (ab16700)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP20] to Vimentin
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
製品の概要
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製品名
Anti-Vimentin antibody [SP20]
Vimentin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [SP20] to Vimentin -
由来種
Rabbit -
特異性
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. -
アプリケーション
適用あり: ICC/IF, WB, IHC-P, Flow Cytmore details -
種交差性
交差種: Human
交差が予測される動物種: Rat, Hamster, Cow, Xenopus laevis -
免疫原
Recombinant full length protein corresponding to Human Vimentin aa 1 to the C-terminus.
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ポジティブ・コントロール
- 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.
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特記事項
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:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.1% Sodium azide
Constituents: 98.9% PBS, 1% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
SP20 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab16700の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | (3) |
1/1000.
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WB | (1) |
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
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IHC-P |
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt |
1/100.
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特記事項 |
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ICC/IF
1/1000. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa. |
IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt
1/100. |
ターゲット情報
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機能
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. -
組織特異性
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. -
関連疾患
Cataract 30 -
配列類似性
Belongs to the intermediate filament family. -
ドメイン
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. -
翻訳後修飾
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. -
細胞内局在
Cytoplasm. - Information by UniProt
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参照データベース
- 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
see all -
製品の状態
Vimentin is found in connective tissue and in the cytoskeleton. -
別名
- CTRCT30 antibody
- Epididymis luminal protein 113 antibody
- FLJ36605 antibody
see all
画像
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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.
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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 kDa why 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.
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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 kDaThis 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.
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Anti-Vimentin antibody [SP20] (ab16700) at 1/100 dilution + HeLa cell lysate
Predicted band size: 53 kDa
Observed band size: 53 kDaThis image was generated using a previous batch manufactured using the hybridoma production method.
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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).
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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.
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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.
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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).
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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (88)
ab16700 は 88 報の論文で使用されています。
- Li W et al. The circ-PITX1 promotes non-small cell lung cancer development via the miR-30e-5p/ITGA6 axis. Cell Cycle 21:304-321 (2022). PubMed: 35007184
- Ukon Y et al. Prostaglandin EP4 Selective Agonist AKDS001 Enhances New Bone Formation by Minimodeling in a Rat Heterotopic Xenograft Model of Human Bone. Front Bioeng Biotechnol 10:845716 (2022). PubMed: 35372320
- Simpson KE et al. Elevated Expression of miR-200c/141 in MDA-MB-231 Cells Suppresses MXRA8 Levels and Impairs Breast Cancer Growth and Metastasis In Vivo. Genes (Basel) 13:N/A (2022). PubMed: 35456497
- Erratico S et al. Effective high-throughput isolation of enriched platelets and circulating pro-angiogenic cells to accelerate skin-wound healing. Cell Mol Life Sci 79:259 (2022). PubMed: 35474498
- Lai IC et al. Selenium Yeast and Fish Oil Combination Diminishes Cancer Stem Cell Traits and Reverses Cisplatin Resistance in A549 Sphere Cells. Nutrients 14:N/A (2022). PubMed: 35956408