MAB3391
Anti-Collagen Type I Antibody, clone 5D8-G9
clone 5D8-G9, Chemicon®, from mouse
Synonim(y):
Anti-CAFYD, Anti-EDSARTH1, Anti-EDSC, Anti-OI1, Anti-OI2, Anti-OI3, Anti-OI4
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About This Item
Kod UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Polecane produkty
pochodzenie biologiczne
mouse
Poziom jakości
forma przeciwciała
purified immunoglobulin
rodzaj przeciwciała
primary antibodies
klon
5D8-G9, monoclonal
reaktywność gatunkowa
pig, canine, sheep, human, bovine
spodziewany brak reakcji z
guinea pig, ground squirrel, rat, kangaroo, horse, chicken, mouse
producent / nazwa handlowa
Chemicon®
metody
ELISA: suitable
flow cytometry: suitable
immunohistochemistry: suitable
western blot: suitable
izotyp
IgG1
numer dostępu NCBI
numer dostępu UniProt
Warunki transportu
wet ice
docelowa modyfikacja potranslacyjna
unmodified
informacje o genach
human ... COL1A1(1277)
Powiązane kategorie
Opis ogólny
Collagens are highly conserved throughout evolution and are characterised by an uninterrupted "Glycine X Y" triplet repeat that is a necessary part of the triple helical structure. Type I collagen (95 kDa) is found in bone, cornea, skin and tendon. Mutations in the encoding gene are associated with osteogenesis imperfecta, Ehlers Danlos syndrome, and idiopathic osteoporosis. Reciprocal translocations between chromosomes 17 and 22, where this gene and the gene for Platelet-derived growth factor beta are located, are associated with a particular type of skin tumor called dermatofibrosarcoma protuberans, resulting from unregulated expression of the growth factor.
Specyficzność
Collagen Type I Antibody is a monoclonal IgG1 antibody which binds an epitope near the C-terminal of Type I Collagen.
Collagen Type I Antibody displays a high affinity for human, bovine and ovine Type I Collagens. There is no evidence for cross reactivity with Collagen Types III, V and VI or connective tissue protein. Antibody reacts only with native, non-denatured Collagen I
Collagen Type I Antibody displays a high affinity for human, bovine and ovine Type I Collagens. There is no evidence for cross reactivity with Collagen Types III, V and VI or connective tissue protein. Antibody reacts only with native, non-denatured Collagen I
Zastosowanie
ELISA: detection of native type I collagen. Capture: 2-4 μg/ml; Detection: A dilution of 10μg/mL of Collagen Type I Antibody will detect 50ng of human Collagen Type I with no reactivity with other Collagens. Higher levels of antibody may be necessary to detect lower concentrations of Collagen. Note antibody will not pair with itself.
Immunohistochemistry:unfixed frozen sections:
Cut 6 μm thick sections from frozen samples using a freezing microtome. Air-dry throughly. Rehydrate with PBS. Blocking is usually not necessary for IF studies, if using DAB, pretreatment for peroxidase is suggested and blocking 2% BSA in PBS will reduce back ground.
Dilute Collagen Type I Antibody in PBS buffer, add to the sample(s) and incubate for 60 minutes. (1:100-1:500).
Wash sections for 10 minutes in PBS, twice. Add FITC conjugated anti-mouse antibody and incubate for 60 minutes.
Wash for 10 minutes in PBS, twice.
Mount sections in glycerol/water (9:1 v/v) containing 1 mM 1,4-phenylenediamine. (for FITC stability or use Chemicon catalog numbers 5096 or 5013.
NOTE:
Preliminary studies suggest that enhanced staining of certain tissues maybe obtained by pretreatment (previous to step 2) of sections with various enzymes (eg 0.1% pepsin in 0.1M HCl, 37°C for 5 minutes).
Flow Cytometry: 1:100
Immunoblotting:1:1000 using NATIVE, non-denatured, non-reduced western blots only. Ramshaw, et.al (1988) "Electrophoretic and electroblotting of native collagens." Anal. Biochem. 168:82-87. Antibody will not work in traditional, reduced western blots.
Briefly, PAGE gels must be prepared for running native collagens.
A 3% (w/v) total acrylamide separating gel, containing 3.2% (w/w) bis-acrylamide as a proportion to a monomer as the crosslinking agent, in 10mM calcium lactate, pH 6.8 is polymerized between vertical glass plates by the addition of 0.05-0.1% TEMED and 0.05-.1% ammonium persulfate (added from a 10% stock solution). The cathode buffer (negative) buffer is 50mM Tris adjusted to pH 6.6 with lactic acid; the anode (positive) buffer is 0.1M lactic acid pH 2.5 {Friis, SJ et al, 1985 J Biochem Biophys Methods 10:301-306.}.
Prior to sample loading the gel is run for at least 90 minutes at 100V. Collagen samples are dissolved at 1mg/mL in either 0.1M lactic acid or 0.1M acetic acid containing 10% sucrose, and 5-10μg per lane is loaded. Methyl green is added as required to assist loading. Electrophoresis is 70V for 5 hours, room temperature.
Blotting uses 0.1M lactic acid pH 2.5, and 20V for 16 hours at room temperature with the collagens migrating toward the cathode.
Immunohistochemistry:unfixed frozen sections:
Cut 6 μm thick sections from frozen samples using a freezing microtome. Air-dry throughly. Rehydrate with PBS. Blocking is usually not necessary for IF studies, if using DAB, pretreatment for peroxidase is suggested and blocking 2% BSA in PBS will reduce back ground.
Dilute Collagen Type I Antibody in PBS buffer, add to the sample(s) and incubate for 60 minutes. (1:100-1:500).
Wash sections for 10 minutes in PBS, twice. Add FITC conjugated anti-mouse antibody and incubate for 60 minutes.
Wash for 10 minutes in PBS, twice.
Mount sections in glycerol/water (9:1 v/v) containing 1 mM 1,4-phenylenediamine. (for FITC stability or use Chemicon catalog numbers 5096 or 5013.
NOTE:
Preliminary studies suggest that enhanced staining of certain tissues maybe obtained by pretreatment (previous to step 2) of sections with various enzymes (eg 0.1% pepsin in 0.1M HCl, 37°C for 5 minutes).
Flow Cytometry: 1:100
Immunoblotting:1:1000 using NATIVE, non-denatured, non-reduced western blots only. Ramshaw, et.al (1988) "Electrophoretic and electroblotting of native collagens." Anal. Biochem. 168:82-87. Antibody will not work in traditional, reduced western blots.
Briefly, PAGE gels must be prepared for running native collagens.
A 3% (w/v) total acrylamide separating gel, containing 3.2% (w/w) bis-acrylamide as a proportion to a monomer as the crosslinking agent, in 10mM calcium lactate, pH 6.8 is polymerized between vertical glass plates by the addition of 0.05-0.1% TEMED and 0.05-.1% ammonium persulfate (added from a 10% stock solution). The cathode buffer (negative) buffer is 50mM Tris adjusted to pH 6.6 with lactic acid; the anode (positive) buffer is 0.1M lactic acid pH 2.5 {Friis, SJ et al, 1985 J Biochem Biophys Methods 10:301-306.}.
Prior to sample loading the gel is run for at least 90 minutes at 100V. Collagen samples are dissolved at 1mg/mL in either 0.1M lactic acid or 0.1M acetic acid containing 10% sucrose, and 5-10μg per lane is loaded. Methyl green is added as required to assist loading. Electrophoresis is 70V for 5 hours, room temperature.
Blotting uses 0.1M lactic acid pH 2.5, and 20V for 16 hours at room temperature with the collagens migrating toward the cathode.
Research Category
Cell Structure
Cell Structure
Research Sub Category
ECM Proteins
ECM Proteins
This Anti-Collagen Type I Antibody, clone 5D8-G9 is validated for use in ELISA, FC, WB, IH for the detection of Collagen Type I.
Postać fizyczna
100μg purified antibody at a concentration of 1mg/mL in 50 mM Tris Acetate, 150 mM NaCl.
The immunoglobulin fraction has been purified by Protein A Chromatography and shown to be >95% pure by coomassie PAGE. The Collagen Type I Antibody is supplied 0.22 micron filtered.
The immunoglobulin fraction has been purified by Protein A Chromatography and shown to be >95% pure by coomassie PAGE. The Collagen Type I Antibody is supplied 0.22 micron filtered.
Format: Purified
Przechowywanie i stabilność
Collagen Type I Antibody is shipped, in liquid form, at ambient temperature. This material is stable for up to 6 months when stored at 2-8°C.
WARNING: The monoclonal reagent solution contains 0.1% sodium azide as a preservative. Due to potential hazards arising from the build up of this material in pipes, spent reagent should be disposed of with liberal volumes of water.
WARNING: The monoclonal reagent solution contains 0.1% sodium azide as a preservative. Due to potential hazards arising from the build up of this material in pipes, spent reagent should be disposed of with liberal volumes of water.
Komentarz do analizy
Control
Human kidney lysate
Human kidney lysate
Informacje prawne
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Oświadczenie o zrzeczeniu się odpowiedzialności
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Kod klasy składowania
12 - Non Combustible Liquids
Klasa zagrożenia wodnego (WGK)
WGK 2
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
Certyfikaty analizy (CoA)
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