Recombinant Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22411-279] to Extracellular matrix protein 1
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-Extracellular matrix protein 1 antibody [EPR22411-279]
See all Extracellular matrix protein 1 primary antibodies -
Description
Rabbit monoclonal [EPR22411-279] to Extracellular matrix protein 1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: 4T1, LLC1, Hepa1-6 whole cell lysate. Mouse and rat skin and liver lysates. Wild-type mouse liver lysate. Flow Cyt (intra): 4T1 cells. IP: Mouse liver and lung whole cell lysate.
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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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22411-279 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab253185 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 |
1/1000. Detects a band of approximately 48, 85 kDa (predicted molecular weight: 63 kDa).
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IP |
1/30.
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IHC-P |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 48, 85 kDa (predicted molecular weight: 63 kDa). |
IP
1/30. |
IHC-P
Use at an assay dependent concentration. |
Target
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Function
Involved in endochondral bone formation as negative regulator of bone mineralization. Stimulates the proliferation of endothelial cells and promotes angiogenesis. Inhibits MMP9 proteolytic activity. -
Tissue specificity
Expressed in breast cancer tissues. Little or no expression observed in normal breast tissues. Expressed in skin; wide expression is observed throughout the dermis with minimal expression in the epidermis. -
Involvement in disease
Defects in ECM1 are the cause of lipoid proteinosis (LiP) [MIM:247100]; also known as lipoid proteinosis of Urbach and Wiethe or hyalinosis cutis et mucosae. LiP is a rare autosomal recessive disorder characterized by generalized thickening of skin, mucosae and certain viscera. Classical features include beaded eyelid papules and laryngeal infiltration leading to hoarseness. Histologically, there is widespread deposition of hyaline material and disruption/reduplication of basement membrane. -
Cellular localization
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 13601 Mouse
- Entrez Gene: 116662 Rat
- SwissProt: Q61508 Mouse
- SwissProt: Q62894 Rat
- Unigene: 3433 Mouse
- Unigene: 97792 Rat
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Alternative names
- ECM 1 antibody
- Ecm1 antibody
- ECM1_HUMAN antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse live tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Extracellular matrix protein 1 with ab253185 at 1/5000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). The section was incubated with ab253185 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer)
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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All lanes : Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185) at 1/1000 dilution
Lane 1 : Wild-type mouse liver lysate
Lane 2 : ECM1 knockout mouse liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 63 kDa
Observed band size: 48,85 kDa why is the actual band size different from the predicted?The wild-type and ECM1 knockout mouse liver lysates were kindly provided by an anonymous collaborator.
ab253185 was shown to specifically react with Extracellular matrix protein 1 in wild-type mouse liver as signal was lost in ECM1 knockout liver. Wild-type and ECM1 knockout samples were subjected to SDS-PAGE. ab253185 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
Exposure time 7.75 secs.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Extracellular matrix protein 1 antibody [EPR22411-279] (ab253185) at 1/1000 dilution
Lane 1 : 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate
Lane 2 : LLC1 (mouse lung carcinoma), whole cell lysate
Lane 3 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate
Lane 4 : Mouse skin lysate
Lane 5 : Mouse liver lysate
Lane 6 : Rat skin lysate
Lane 7 : Rat liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 63 kDa
Observed band size: 48,85 kDa why is the actual band size different from the predicted?Mouse ECM1 has two isoforms: A long isoform reported to be 85 kDa, and the shorter form 48 kDa.
The molecular weight observed is consistent with what has been described in the literature (PMID: 7608209; PMID: 23202415).
Exposure times: Lanes 1-2: 7.75 seconds; Lane 3: 1 second; Lanes 4-7: 10 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Extracellular matrix protein 1 was immunoprecipitated from 0.35 mg mouse lung lysate with ab253185 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253185 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse lung lysate 10µg.
Lane 2: ab253185 IP in mouse lung lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253185 in mouse lung lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Extracellular matrix protein 1 was immunoprecipitated from 0.35 mg mouse liver lysate with ab253185 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253185 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse liver lysate 10µg.
Lane 2: ab253185 IP in mouse liver lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253185 in mouse liver lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 mins.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized 4T1 (mouse mammary gland carcinoma epithelial cell) cells labeling Extracellular matrix protein 1 with ab253185 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). A 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 (1)
ab253185 has been referenced in 1 publication.
- Liu Y et al. MiR-122-5p promotes peritoneal fibrosis in a rat model of peritoneal dialysis by targeting Smad5 to activate Wnt/β-catenin pathway. Ren Fail 44:191-203 (2022). PubMed: 35170385