重组Anti-TIMP1抗体[EPR18352] (ab211926)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18352] to TIMP1
- Suitable for: IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-TIMP1抗体[EPR18352]
参阅全部 TIMP1 一抗 -
描述
兔单克隆抗体[EPR18352] to TIMP1 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human prostate cancer lysate; HeLa, SK-OV-3, HT-29, and U-87 MG whole cell lysates. IHC-P: Human colon, pancreas, lung cancer, medullary thyroid carcinoma and prostate cancer tissues. ICC/IF: SK-OV-3 and HT-29 cells.
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常规说明
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18352 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab211926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。
应用 | Ab评论 | 说明 |
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IHC-P | (3) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
WB | (1) |
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).
|
ICC/IF | (3) |
1/500.
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说明 |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa). |
ICC/IF
1/500. |
靶标
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功能
Complexes with metalloproteinases (such as collagenases) and irreversibly inactivates them by binding to their catalytic zinc cofactor. Also mediates erythropoiesis in vitro; but, unlike IL-3, it is species-specific, stimulating the growth and differentiation of only human and murine erythroid progenitors. Known to act on MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13 and MMP-16. Does not act on MMP-14. -
序列相似性
Belongs to the protease inhibitor I35 (TIMP) family.
Contains 1 NTR domain. -
翻译后修饰
The activity of TIMP1 is dependent on the presence of disulfide bonds. -
细胞定位
Secreted. - Information by UniProt
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数据库链接
- Entrez Gene: 7076 Human
- Omim: 305370 Human
- SwissProt: P01033 Human
- Unigene: 522632 Human
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别名
- Clgi antibody
- Collagenase inhibitor antibody
- Collagenase inhibitor, Human antibody
see all
图片
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All lanes : Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TIMP1 knockout HeLa cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : SH-SY5Y cell lysate
Lane 5 : U-87 MG cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab211926 was shown to bind specifically to TIMP1. A band was observed at 30 kDa in wild-type HeLa cell lysates with no signal observed at this size in TIMP1 knockout cell line ab264022 (knockout cell lysate ab260091). The identity of bands observed at higher molecular weights has not been determined. To generate this image, wild-type and TIMP1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TIMP1 knockout HeLa cell lysate
Lane 3 : HT-1080 treated with 200ng/ml 12-O Tetradecanoylphorbol-13-acetate (TPA) for 24 hours, cell lysate
Lane 4 : Untreated HT-1080 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab211926 was shown to bind specifically to TIMP1. A band was observed at 30 kDa in wild-type HeLa cell lysates with no signal observed at this size in TIMP1 knockout cell line ab264022 (knockout cell lysate ab260091). The identity of bands observed at higher molecular weights has not been determined. To generate this image, wild-type and TIMP1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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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Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human lung cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution + Human prostate cancer lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16517973).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (Human ovarian cancer cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SK-OV-3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
All lanes : Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution
Lane 1 : A549 Vehicle Control BFA (0 u/mL, 6 h) cell lysate
Lane 2 : A549 Treated BFA (5 u/mL, 6 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in black. In Western blot, ab211926 was shown to bind specifically to TIMP1. 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 before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) at 1/50000 dilution and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution
Lane 1 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 2 : SK-OV-3 (Human ovarian cancer cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Binding in these cell lines was extremely weak and requires optimization.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human colon neuroendocrine cell is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (Human colorectal adenocarcinoma cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HT-29 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human islet is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded human medullary thyroid carcinoma tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human medullary thyroid carcinoma is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human prostate cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (30)
ab211926 被引用在 30 文献中.
- Li YK et al. Validation of ESM1 Related to Ovarian Cancer and the Biological Function and Prognostic Significance. Int J Biol Sci 19:258-280 (2023). PubMed: 36594088
- Fang F & Yuan Q Anlotinib inhibits the proliferation, migration and invasion, and induces apoptosis of breast cancer cells by downregulating TFAP2C. Oncol Lett 23:46 (2022). PubMed: 34976158
- Chu Z et al. Betulonic Acid, as One of the Active Components of the Celastrus orbiculatus Extract, Inhibits the Invasion and Metastasis of Gastric Cancer Cells by Mediating Cytoskeleton Rearrangement In Vitro. Molecules 27:N/A (2022). PubMed: 35164287
- Wang LJ et al. Tumor-associated macrophages related signature in glioma. Aging (Albany NY) 14:2720-2735 (2022). PubMed: 35332109
- Wang H et al. Poria Acid, Triterpenoids Extracted from Poria cocos, Inhibits the Invasion and Metastasis of Gastric Cancer Cells. Molecules 27:N/A (2022). PubMed: 35684565