Collagen Type I 抗体
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- Key Features
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- High quality Collagen Type I Primary Antibody for the detection of COL1.
- Reliable product with high standard validation data
- 抗原 See all Collagen Type I (COL1) 抗体
- Collagen Type I (COL1) (Collagen, Type I (COL1))
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适用
- 人, 大鼠, 小鼠, Cow, 哺乳动物
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This Collagen Type I antibody is un-conjugated
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应用范围
- Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Immunoprecipitation (IP)
- 交叉反应 (详细)
- Typically negligible cross-reactivity against other types of collagens was detected by ELISA against purified standards. Some class-specific anti-collagens may be specific for three-dimensional epitopes which may result in diminished reactivity with denatured collagen or formalin-fixed, paraffin embedded tissues. This antibody reacts with most mammalian Type I collagens and has negligible cross-reactivity with Type II, III, IV, V or VI collagens. Non-specific cross-reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible.
- 纯化方法
- COLLAGEN I Antibody has been prepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against other collagens, human serum proteins and non-collagen extracellular matrix proteins to remove any unwanted specificities.
- 免疫原
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Immunogen: Collagen Type I from human and bovine placenta
Immunogen Type: Native Protein
- 亚型
- IgG
- Product Specific Information
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- What can the Collagen Type I Antibody ABIN5596819 be used for?
- This polyclonal Collagen Type I Antibody detects Collagen Type I. The Collagen Type I Antibody has been validated for various applications and can be used for the detection of Collagen Type I and derivatives by Western Blotting, ELISA, Immunohistochemistry, Immunoprecipitation.
- What validation data is available for this Collagen Type I Antibody?
- The Primary Antibody is referenced in 62 publications and is characterized by a proven, very high reliability. It has currently 41 product images that show its performance in a variety of applications. The product is currently available in 100 μg, 500 μg quantities. Collagen Type I Antibody for the detection of Collagen Type I and derivatives.
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- 应用备注
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Immunohistochemistry Dilution: 1:50 - 1:200
Application Note: Anti-Collagen antibodies have been used for indirect trapping ELISA for quantitation of antigen in serum using a standard curve, for immunoprecipitation and for native (non-denaturing, non-dissociating) PAGE and western blotting for highly sensitive qualitative analysis.
Western Blot Dilution: 1:1,000 - 1:10,000
Immunoprecipitation Dilution: 1:100
ELISA Dilution: 1:5,000 - 1:50,000
- 限制
- 仅限研究用
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- by
- NanoTemper Technologies
- No.
- #103813
- 日期
- 2019.07.23
- 抗原
- COL1
- Lot Number
- 41897
- Method validated
- Unfolding Profile
- Positive Control
- ABIN5596819
- Negative Control
- Notes
Passed. ABIN5596819 showed Ti at 79.6°C and a clear unfolding profile with one unfolding event. This suggests that the antibody is properly folded and functional.
- Primary Antibody
- Secondary Antibody
- Full Protocol
- Dilute ABIN5596819 in PBS buffer (Roth, 1058.1, lot 285231988) to get a final volume of 30µl at a concentration of 0.1µM.
- Load sample into Tycho capillary (NanoTemper Technologies, TY-C001).
- Run Tycho measurement.
- Experimental Notes
Tycho is designed to run quick and precise protein quality check experiments. Tycho uses intrinsic protein fluorescence to follow protein unfolding while running a fast thermal ramp, yielding results in 3min. A protein’s unfolding behavior is characterized by various parameters, most notably the inflection temperature (Ti). The Ti can be used to identify properly folded protein, to compare different batches, or to analyze the influence of storage/transport conditions on a protein. An absence of Ti would suggest that the protein is already unfolded and therefore most likely nonfunctional.
生效 #103813 (Unfolding Profile)Validation ImagesFull Methods -
- by
- MS Validated Antibodies
- No.
- #300048
- 日期
- 2023.08.07
- 抗原
- COL1
- Lot Number
- 47973
- Method validated
- Immunohistochemistry
- Positive Control
Human TMA
Monoclonal rabbit anti-human COL1A2 monoclonal antibody (HL2048)
- Negative Control
- Notes
Passed. Staining of collagen type I using ABIN5596819 is consistent with the expected staining pattern.
- Primary Antibody
- ABIN5596819
- Secondary Antibody
- EnVision Polymer-HRP mouse/rabbit Kit, Dako REAL, K5007
- Full Protocol
- Slide preparation
- Mount 2,5 µm FFPE tissue sections on superfrost slides.
- Deparaffinize tissue sections 3x 5 min in xylene.
- Rehydrate tissue sections in a descending ethanol series for 1 min each 100%, 96%, and 80% ethanol.
- Rinse tissue sections for 5 min in TBST buffer (DAKO, K8000).
- Epitope retrieval
- Autoclave tissue sections for 5 min at 121 °C in 1x Tris-EDTA-citrate buffer pH7.8 (20x Tris-EDTA-citrate buffer stock solution: 5 g Trizma base (Sigma-Aldrich, T1503), 10 g EDTA (Merck, 1.08418), 6.4g tri-sodium citrate (Sigma-Aldrich, C0909), adjust to pH 7.8 using HCL 1 M, ad 1 L with dH2O).
- Rinse tissue sections for 5 min in TBST buffer.
- Peroxidase blocking
- Incubate tissue sections for 10 min in Peroxidase-Blocking Solution (Dako REAL, S2023).
- Rinse tissue sections 2x for 5 min in TBST buffer.
- Antibody incubation
- Dilute primary rabbit anti-collagen type I antibody (antibodies-online, ABIN5596819, lot 47973) diluted 1:15 or 1:50 in antibody diluent (Dako REAL, S2022).
- Cover tissue section with 100-200 µl diluted antibody.
- Incubate tissue sections for 1 h at 37 °C in a moist chamber.
- Rinse tissue sections for 5 min in TBST buffer.
- Apply EnVision Polymer-HRP mouse/rabbit Kit (Dako REAL, K5007) according to manufacturer’s recommendation.
- Rinse tissue sections 2x for 5 min in TBST buffer.
- Staining
- Cover slides for 10 min with DAB-Chromogen (EnVision Polymer-HRP mouse/rabbit Kit, Dako REAL, K5007).
- Wash slides thoroughly with dH2O.
- Counterstain for 15 sec with Hematoxylin (Mayers Hematoxylin: 200ml ddH2O, 0,2g Hematoxylin (Serva, 24420.02), 10 g aluminium potassium sulfate dodecahydrate (Merck, 1.01047), 0,04 g sodium iodate (Merck, 1.06525), 10 g chloral hydrate (Sigma-Aldrich, 15307)).
- Develop for 15 sec in H2O.
- Dehydrate tissue sections in an ascending ethanol series for 1 min each 80%, 96%, 100% ethanol.
- Wash tissue sectiona 3x 5 min in xylene.
- Apply mounting medium and coverslips.
- Image acquisition
- o Acquire images using a Galileo TMAtic (ISENET).
- Experimental Notes
For antibody comparison an antibody test TMA was used that contained 80 normal tissues from 21 different organs and 95 neoplastic tissues from 18 different tumor types. For ABIN5596819 the staining was typically fibrillar. Positive fibrillar structures were often located adjacent to benign and malignant epithelial structures, in smooth muscles or around vessels of all sizes.
The anti-collagen type I antibody ABIN5596819 shows a predominantly fibrillar staining pattern involving stroma components that are often located adjacent to benign and malignant epithelial structures, in smooth muscle or around vessels of all sizes. Collagen I staining is also seen in the stroma of many tumors. The staining of these fibrillar structures by ABIN5596819 is often faint at a dilution of 1:50. Staining of fibrils by ABIN5596819 is stronger at a dilution of 1:15 but in this case a significant background staining occurs in many epithelial tissues. Of note, a membranous staining of intratubular testicular cells was seen by ABIN5596819. Considering the expected staining pattern of an anti-collagen type I antibody, this may represent a cross-reactivity of ABIN5596819.
According to the rather ubiquitous nature of collagen I expression, orthogonal validation is not optimally suited for the validation of collagen I antibodies. In agreement with RNA screening studies, including the Human Protein Atlas (HPA) RNA-seq tissue dataset, the FANTOM5 project, and the Genotype-Tissue Expression (GTEx) project (all summarized in https://www.proteinatlas.org/ENSG00000108821-COL1A1/tissue), collagen I staining by ABIN5596819 was significant in the placenta, smooth muscle, urinary bladder and the endometrium. These are the tissues with the highest recorded RNA expression among the tissues analyzed in this project.
生效 #300048 (Immunohistochemistry)Validation ImagesFull Methods -
- 状态
- Liquid
- 浓度
- 1.0 mg/mL
- 缓冲液
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Buffer: 0.125 M Sodium Borate, 0.075 M Sodium Chloride, 0.005 M EDTA, pH 8.0
Stabilizer: None
- 储存液
- Sodium azide
- 注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 储存条件
- 4 °C,-20 °C
- 储存方法
- Store vial at 4° C prior to opening. This product is stable at 4° C as an undiluted liquid. Dilute only prior to immediate use. For extended storage, mix with an equal volume of glycerol, aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing.
- 有效期
- 12 months
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A Scaffold-Free 3-D Co-Culture Mimics the Major Features of the Reverse Warburg Effect In Vitro." in: Cells, Vol. 9, Issue 8, (2020) (PubMed).
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- 抗原
- Collagen Type I (COL1) (Collagen, Type I (COL1))
- 别名
- COLLAGEN Type I (COL1 产品)
- 别名
- collagen type III alpha 1 chain antibody, COL3A1 antibody
- 背景
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Synonyms: Collagen Of Skin Tendon And Bone antibody, Collagen Type 1 antibody, Collagen type I alpha 1 antibody, Collagen type I alpha 2 antibody, OI4 antibody, Osteogenesis Imperfecta Type IV antibody
Background: Collagens are highly conserved throughout evolution and are characterized by an uninterrupted ''Glycine-X-Y'' triplet repeat that is a necessary part of the triple helical structure. For these reasons, it is often extremely difficult to generate antibodies with specificities to collagens. The development of 'type' specific antibodies is dependent on NON-DENATURED three-dimensional epitopes. Rockland extensively purifies collagens for immunization from human and bovine placenta and cartilage by limited pepsin digestion and selective salt precipitation. This preparation results in a native conformation of the protein. Antibodies are isolated from rabbit antiserum and are extensively cross-adsorbed by immunoaffinity purification to produce 'type' specific antibodies. Greatly diminished reactivity and selectivity of these antibodies will result if denaturing and reducing conditions are used for SDS-PAGE and immunoblotting. Ideal for investigators involved in Cell Biology, Signal Transduction and Stem Cell research.
Gene Name: COL1A1
- 基因ID
- 1277
- UniProt
- P02452
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