重组Anti-Collagen VI抗体[EPR17072] (ab182744)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17072] to Collagen VI
- Suitable for: WB, ICC/IF, IHC-P, mIHC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Collagen VI抗体[EPR17072]
参阅全部 Collagen VI 一抗 -
描述
兔单克隆抗体[EPR17072] to Collagen VI -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IHC-P, mIHCmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human skeletal muscle, Human placenta, Human fetal brain, Human fetal heart, Human fetal kidney, Human fetal spleen, Mouse heart, Mouse kidney, Mouse spleen, Rat kidney and Rat spleen lysates; HEK293T, WI-38 and NIH/3T3 whole cell lysates. IHC-P: Human liver, Human cardiac muscle, Mouse kidney and Rat stomach tissues. mIHC: Human liver and Human prostate gland tissues, human breast. ICC/IF: HeLa 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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR17072 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-Collagen VI antibody [EPR17072] (ab200429)
- Alexa Fluor® 647 Anti-Collagen VI antibody [EPR17072] (ab200430)
- HRP Anti-Collagen VI antibody [EPR17072] (ab200596)
- Alexa Fluor® 594 Anti-Collagen VI antibody [EPR17072] (ab207292)
- Anti-Collagen VI antibody [EPR17072] - Low endotoxin, Azide free (ab229450)
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab182744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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WB | (1) |
1/2000. Detects a band of approximately 147 kDa (predicted molecular weight: 109 kDa).
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ICC/IF |
1/200.
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IHC-P | (3) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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mIHC |
1/500 - 1/1000.
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说明 |
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WB
1/2000. Detects a band of approximately 147 kDa (predicted molecular weight: 109 kDa). |
ICC/IF
1/200. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
mIHC
1/500 - 1/1000. |
靶标
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功能
Collagen VI acts as a cell-binding protein. -
疾病相关
Defects in COL6A1 are a cause of Bethlem myopathy (BM) [MIM:158810]. BM is a rare autosomal dominant proximal myopathy characterized by early childhood onset (complete penetrance by the age of 5) and joint contractures most frequently affecting the elbows and ankles.
Defects in COL6A1 are a cause of Ullrich congenital muscular dystrophy (UCMD) [MIM:254090]; also known as Ullrich scleroatonic muscular dystrophy. UCMD is an autosomal recessive congenital myopathy characterized by muscle weakness and multiple joint contractures, generally noted at birth or early infancy. The clinical course is more severe than in Bethlem myopathy. -
序列相似性
Belongs to the type VI collagen family.
Contains 3 VWFA domains. -
翻译后修饰
Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains. -
细胞定位
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 1291 Human
- Entrez Gene: 1292 Human
- Entrez Gene: 1293 Human
- Entrez Gene: 12833 Mouse
- Entrez Gene: 12834 Mouse
- Entrez Gene: 12835 Mouse
- Entrez Gene: 294337 Rat
- Entrez Gene: 361821 Rat
see all -
别名
- Alpha 1 (VI) chain (61 AA) antibody
- CO6A1_HUMAN antibody
- COL6A1 antibody
see all
图片
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This data was developed using the same antibody clone in a different buffer formulation (ab271938).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast labelling Cytokeratin 18 with ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with ab236439 at 1/2000 dilution ( 0.519 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Panel A: merged staining of anti-Cytokeratin 14 (magenta; Opal™690), anti-Cytokeratin 18 (green; Opal™520) and anti-Collagen VI (red; Opal™570) on human breast.
Panel B: anti-Cytokeratin 18 stained on luminal epithelial cells.
Panel C: anti-Collagen VI stained on stroma.
Panel D: anti-Cytokeratin 14 stained on myoepithelial cells.The section was incubated in three rounds of staining: in the order of ab236439 for 30 mins, ab93741 for 10 mins, and ab271938 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
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Fluorescence multiplex immunohistochemical analysis of human adrenal gland (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-PCSK2 stained on adrenal medulla (ab274418; gray; Opal™690) at 1:2000 (0.263 μg/ml) [Panel B], anti-CYP11A1 stained on adrenal cortex (ab272494; green; Opal™520) at 1:10000 (0.053 μg/ml) [Panel C], and anti-Collagen VI stained on extracellular matrix (ab182744; red; Opal™570) at 1:1000 (1.518 μg/ml) [Panel D] on human adrenal gland. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab274418, ab272494, and ab182744 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
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Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-Collagen VI (ab271938, magenta; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Prostate Specific Antigen (ab76113, red; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on luminal cells. Panel C: anti-Cytokeratin 5 stained on basal cells. Panel D: anti-Collagen VI stained on stroma. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab271938 (1/500), ab236216 (1/400), and ab76113 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.
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Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around sinusoidal endothelial basement membranes is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : COL6A1 knockout HEK293T cell lysate
Lane 3 : Human skeletal muscle tissue 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: 109 kDa
Observed band size: 136 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab182744 observed at 136 kDa. Red - loading control ab8245 observed at 36 kDa.
ab182744 Anti-Collagen VI antibody [EPR17072] was shown to specifically react with Collagen VI antibody in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265060 (knockout cell lysate ab256879) was used. Wild-type and Collagen VI antibody knockout samples were subjected to SDS-PAGE. ab182744 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 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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All lanes : Anti-Collagen VI antibody [EPR17072] (ab182744)
Lane 1 : Wild-type HEK293 whole cell lysate
Lane 2 : COL6A1 knockout HEK293 whole cell lysate
Lane 3 : Human Skeletal Muscle whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 109 kDaLanes 1 - 3: Merged signal (red and green). Green - ab182744 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab182744 was shown to recognize Collagen VI in wild-type HEK293 cells as signal was lost at the expected MW in COL6A1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and COL6A1 knockout samples were subjected to SDS-PAGE. Ab182744 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue.
Panel A: Merged staining of Collagen VI (ab182744; green), anti-CD68 (ab213363; red) and anti-Lamin B1 (ab229025; magenta).
Panel B: Anti-Collagen VI (green) stained on extracellular matrix.
Panel C: Anti-CD68 (red) stained on Kupffer cells.
Panel D: Anti-Lamin B1 (magenta) stained on nuclear envelope.
Key protocol steps: The section was incubated in three rounds of staining with ab182744 (1/1000 dilution), ab213363 (1/1000 dilution) and ab229025 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
DAPI was used as a nuclear counter stain. A ready-to-use anti-Rabbit and Mouse Polymer HRP was used as a secondary.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Collagen VI with ab182744 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control1 - ab182744 at 1/200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining on Human cardiac sarcolemma and interstitium is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around basement membranes of Mouse renal tubules is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Collagen VI with ab182744 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Positive staining around Rat gastric epithelial basement membranes is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/20000 dilution
Lane 1 : Human skeletal muscle lysate
Lane 2 : WI-38 (Human fetal lung fibroblast cells) whole cell lysate
Lane 3 : Human placenta lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 109 kDa
Observed band size: 147 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Due to posttranslational modifications, observed MW is greater than the predicted MW.
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All lanes : Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/2000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 109 kDa
Observed band size: 147 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Due to posttranslational modifications, observed MW is greater than the predicted MW.
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All lanes : Anti-Collagen VI antibody [EPR17072] (ab182744) at 1/2000 dilution
Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat kidney lysate
Lane 5 : Rat spleen lysate
Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 109 kDa
Observed band size: 147 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Due to posttranslational modifications, observed MW is greater than the predicted MW.
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)
ab182744 被引用在 30 文献中.
- Malerba A et al. Systemic Delivery of a Monoclonal Antibody to Immunologically Block Myostatin in the A17 Mouse Model of OPMD. Methods Mol Biol 2587:557-568 (2023). PubMed: 36401050
- Wu C et al. Assessment of stromal SCD-induced drug resistance of PDAC using 3D-printed zPDX model chips. iScience 26:105723 (2023). PubMed: 36590169
- Sterenczak KA et al. The Antibiotic Kitasamycin-A Potential Agent for Specific Fibrosis Preventing Therapy after Fistulating Glaucoma Surgery? Pharmaceutics 15:N/A (2023). PubMed: 36839651
- Lu L et al. Anaplastic transformation in thyroid cancer revealed by single-cell transcriptomics. J Clin Invest 133:N/A (2023). PubMed: 37053016
- Koop K et al. Interleukin 36 receptor-inducible matrix metalloproteinase 13 mediates intestinal fibrosis. Front Immunol 14:1163198 (2023). PubMed: 37207229