Recombinant Anti-Collagen I antibody [EPR22894-89] (ab260043)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22894-89] to Collagen I
- Suitable for: Flow Cyt (Intra), WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Collagen I antibody [EPR22894-89]
See all Collagen I primary antibodies -
Description
Rabbit monoclonal [EPR22894-89] to Collagen I -
Host species
Rabbit -
Specificity
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IPmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NIH/3T3 culture supernatant, Rat lung organ culture supernatant. Human lung tissue lysate, HFF-1 whole cell lysate and culture supernatant, A549, MDA-MB-231, A431 and SW480 whole cell lysates and MDA-MB-231 supernatant lysate. IP: Mouse lung lysate. Flow Cyt (intra): HFF-1 and NIH/3T3 cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22894-89 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab260043 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/500.
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WB | (1) |
1/1000. Predicted molecular weight: 139 kDa.
Positive Control: Hu stomach, skin and adrenal gland tissue lysates. Acid or enzyme treatment with pepsin is a better method to isolate collagen. Continuous refrigeration throughout collagen extraction is important to avoid degradation and denaturation. Take care with pH, temperature, and concentration to avoid collagen polymerization. |
IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 139 kDa. Positive Control: Hu stomach, skin and adrenal gland tissue lysates. Acid or enzyme treatment with pepsin is a better method to isolate collagen. Continuous refrigeration throughout collagen extraction is important to avoid degradation and denaturation. Take care with pH, temperature, and concentration to avoid collagen polymerization. |
IP
1/30. |
Target
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Function
Type I collagen is a member of group I collagen (fibrillar forming collagen). -
Tissue specificity
Forms the fibrils of tendon, ligaments and bones. In bones the fibrils are mineralized with calcium hydroxyapatite. -
Involvement in disease
Defects in COL1A1 are the cause of Caffey disease (CAFFD) [MIM:114000]; also known as infantile cortical hyperostosis. Caffey disease is characterized by an infantile episode of massive subperiosteal new bone formation that typically involves the diaphyses of the long bones, mandible, and clavicles. The involved bones may also appear inflamed, with painful swelling and systemic fever often accompanying the illness. The bone changes usually begin before 5 months of age and resolve before 2 years of age.
Defects in COL1A1 are a cause of Ehlers-Danlos syndrome type 1 (EDS1) [MIM:130000]; also known as Ehlers-Danlos syndrome gravis. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS1 is the severe form of classic Ehlers-Danlos syndrome.
Defects in COL1A1 are the cause of Ehlers-Danlos syndrome type 7A (EDS7A) [MIM:130060]; also known as autosomal dominant Ehlers-Danlos syndrome type VII. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS7A is marked by bilateral congenital hip dislocation, hyperlaxity of the joints, and recurrent partial dislocations.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 1 (OI1) [MIM:166200]. A dominantly inherited connective tissue disorder characterized by bone fragility and blue sclerae. Osteogenesis imperfecta type 1 is non-deforming with normal height or mild short stature, and no dentinogenesis imperfecta.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 2A (OI2A) [MIM:166210]; also known as osteogenesis imperfecta congenita. A connective tissue disorder characterized by bone fragility, with many perinatal fractures, severe bowing of long bones, undermineralization, and death in the perinatal period due to respiratory insufficiency.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 3 (OI3) [MIM:259420]. A connective tissue disorder characterized by progressively deforming bones, very short stature, a triangular face, severe scoliosis, grayish sclera, and dentinogenesis imperfecta.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 4 (OI4) [MIM:166220]; also known as osteogenesis imperfecta with normal sclerae. A connective tissue disorder characterized by moderately short stature, mild to moderate scoliosis, grayish or white sclera and dentinogenesis imperfecta.
Genetic variations in COL1A1 are a cause of susceptibility to osteoporosis (OSTEOP) [MIM:166710]; also known as involutional or senile osteoporosis or postmenopausal osteoporosis. Osteoporosis is characterized by reduced bone mass, disruption of bone microarchitecture without alteration in the composition of bone. Osteoporotic bones are more at risk of fracture.
Note=A chromosomal aberration involving COL1A1 is found in dermatofibrosarcoma protuberans. Translocation t(17;22)(q22;q13) with PDGF. -
Sequence similarities
Belongs to the fibrillar collagen family.
Contains 1 fibrillar collagen NC1 domain.
Contains 1 VWFC domain. -
Post-translational
modificationsProline residues at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains. Proline residues at the second position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some of the chains.
O-linked glycan consists of a Glc-Gal disaccharide bound to the oxygen atom of a post-translationally added hydroxyl group. -
Cellular localization
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 1277 Human
- Entrez Gene: 1278 Human
- Entrez Gene: 12842 Mouse
- Entrez Gene: 12843 Mouse
- Entrez Gene: 29393 Rat
- Entrez Gene: 84352 Rat
- Omim: 120150 Human
- Omim: 120160 Human
see all -
Alternative names
- Alpha 1 type I collagen antibody
- Alpha 2 type I collagen antibody
- alpha 2 type I procollagen antibody
see all
Images
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All lanes : ab260043 and ab255809 at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : MDA-MB-231(Human breast adenocarcinoma epithelial cell) supernatant lysate
Lane 4 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 5 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 139 kDa
Observed band size: 220,35 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Exposure Time: ab260043: 20 seconds, ab255809: 180 seconds.
ab181602 was used as loading control.
Compared with ab260043, ab255809 has lower background. We recommend ab255809 as an alternative for western blotting.
For better use ab255809, we recommend loading higher amount of lysate or using lower antibody dilution.
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All lanes : Anti-Collagen I antibody [EPR22894-89] (ab260043) at 1/5000 dilution
Lane 1 : HFF-1 (human skin fibroblast) whole cell lysate at 20 µg
Lane 2 : HFF-1 culture supernatant at 10 µl
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/25000 dilution
Predicted band size: 139 kDa
Observed band size: 220,35 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Procollagen I (220 kDa) and cleaved C-propeptide (35 kDa) are observed.
The molecular weight observed is consistent with what has been described in the literature (PMID: 23940311).
Exposure time: Lane 1: 1 second; Lane 2: 3 seconds.
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All lanes : Anti-Collagen I antibody [EPR22894-89] (ab260043) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryonic fibroblast) culture supernatant at 10 µl
Lane 2 : Rat lung organ culture supernatant at 10 µl
Lane 3 : Human lung tissue lysate at 10 µg
Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/25000 dilution
Lane 3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 139 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Cleaved collagen I C-propeptide (35 kDa) is observed.
The molecular weight observed is consistent with what has been described in the literature (PMID: 23940311).
Exposure time: Lanes 1-2: 3 seconds; Lane 3: 136 seconds.
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Collagen I was immunoprecipitated from 0.35 mg mouse lung lysate 10µg with ab260043 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260043 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Lane 1: Mouse lung lysate 10µg
Lane 2: ab260043 IP in mouse lung lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab260043 in mouse lung lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Collagen I with ab260043 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HFF-1 (Human skin fibroblast) cells labeling Collagen I with ab260043 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (60)
ab260043 has been referenced in 60 publications.
- Liu S et al. Immunomodulatory hybrid micro-nanofiber scaffolds enhance vascular regeneration. Bioact Mater 21:464-482 (2023). PubMed: 36185748
- Guo Z et al. β-Klotho Promotes the Development of Intrauterine Adhesions via the PI3K/AKT Signaling Pathway. Int J Mol Sci 23:N/A (2022). PubMed: 36232594
- Wu C et al. Tannic acid-loaded hydrogel coating endues polypropylene mesh with hemostatic and anti-inflammatory capacity for facilitating pelvic floor repair. Regen Biomater 9:rbac074 (2022). PubMed: 36267152
- Gerwinn T et al. Improved contractile potential in detrusor microtissues from pediatric patients with end stage lower urinary tract dysfunction. Front Cell Dev Biol 10:1007265 (2022). PubMed: 36268506
- Wang Q et al. miR-153-3p inhibits osteogenic differentiation of BMSCs by down-regulating the expression of RUNX2 in a high glucose environment. Am J Transl Res 14:7027-7039 (2022). PubMed: 36398274