Prolyl 4-hydroxylase subunit beta - AF0910-1

Product Description for Prolyl 4-hydroxylase subunit beta

Mouse anti Human Prolyl 4-hydroxylase subunit beta 3-2B12.
Presentation: Purified
Product is tested for Frozen Sections, Western blot / Immunoblot, Paraffin Sections.

Properties for Prolyl 4-hydroxylase subunit beta

Product Category	Primary Antibodies
Quantity	0.1 mg
Synonyms	Fibroblast marker, P4HB, Prolyl 4-hydroxylase beta
Presentation	Purified
Reactivity	Hu
Applications	C, P, WB
Clonality	Monoclonal
Clone	3-2B12
Host	Mouse
Isotype	IgG1
Shipping to	Worldwide
PDF datasheet	View Datasheet
Manufacturer	OriGene Technologies GmbH
Material safety datasheet	MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen	Immunogen: Native Human merosin M-chain. The epitope has not been further determined.
Isotype control	AM03095PU-N, SM10P (for use in human samples)
Add. information	Has been reported to cross react with rat P4H beta. Cross reativity to rat has been observed in Western blot, IHC and ELISA. Some users have reported cross reactivities with myoepithelial and urothelial cells.
Application	Immunohistochemistry on Frozen Sections: 0.25-0.5 µg/ml (1/400 - 1/800). Immunohistochemistry on Paraffin Sections: 4 µg/ml (1/50). Microwave pretreatment for antigen retrieval is recommended. Suggested Positive Control: Human skin, tonsil. Has been described to work in Western Blots (Ref.1) showing a band at approx. 60kDa.
Background	Collagen prolyl 4-hydroxylases (P4Hs) play an essential role in the synthesis of all collagens. Two alpha and two beta subunits assemble into P4H tetramers in which protein disulfide isomerase (PDI) acts as the beta subunit. The three dimensional structure of many extracellular proteins is stabilized by the formation of disulphide bonds. Studies suggest that a microsomal enzyme known as Protein Disulphide Isomerase (PDI) is involved in disulphide-bond formation and isomerization, as well as the reduction of disulphide bonds in proteins. PDI, which catalyses disulphide interchange between thiols and protein dilsulphides, has also been referred to as thiol:protein-disulphide oxidoreductase and as glutathione:insulin transhydrogenase because of its role in reduction of disulphide bonds. The highly conserved sequence Lys-Asp-Glu-Leu (KDEL) is present at the carboxy-terminus of PDI and other soluble endoplasmic reticulum (ER) resident proteins including the 78 and 94 kDa glucose regulated proteins (GRP78 and GRP94 respectively). The presence of carboxy-terminal KDEL appears to be necessary for ER retention and appears to be sufficient to reduce the secretion of proteins from the ER. This retention is reported to be mediated by a KDEL receptor.
Concentration	0.2 mg/ml (after reconstitution)
Protocols	Protocol for Frozen Sections: Incubations are done at RT. Water is of double distilled or comparable quality. 1. Fix fresh frozen sections in ice-cold acetone for 10 min 2. Block endogenous peroxidase with 100ml 0.15M sodium azide / 0.15% H₂O₂ in PBS - Wash in PBS 3. Block with 10% Normal Rabbit Serum for 30 min in a humid chamber 4. Incubate with primary antibody Cat.-No AF0910-1 for 1 hour in a humid chamber - Wash in PBS 5. Incubate with secondary antibody (peroxidase-conjugated Rabbit Anti Mouse IgG Cat.-No R1253HRP), at the recommended dilution for 1 hour in a humid chamber - Wash in PBS 6. Incubate with AEC substrate (3-Amino-9-Ethylcarbazol) for 12 minutes - Wash in PBS 7. Counterstain with Mayer's hemalum. Protocol for Paraffin Sections: Incubations are done at RT. Water is of double distilled or comparable quality. 1. Rehydrate paraffin sections 2. Put the slides in a cuvette with 250 ml 0.01 M citrate buffer pH 6.0 3. Heat the slides in a microwave oven for 2 x 7 min and 700 Watt 4. Leave the slides in the buffer for 20 min 5. Block endogenous peroxidase with 1% H₂O₂ in water - Wash in PBS 6. Block with 10% Normal Goat Serum for 30 min in a humid chamber 7. Incubate with primary antibody cat.-No AF0910-1 for 1 hour in a humid chamber - Wash in PBS 8. Incubate with secondary antibody ( peroxidase-conjugated Goat Anti Mouse IgG (H+L), at a dilution recommended by the manufacturer for 1 hour in a humid chamber - Wash in PBS 9. Incubate with AEC substrate (3-Amino-9-Ethylcarbazol) for 12 minutes - Wash in PBS 10. Counterstain with Mayer's hemalum.
Product Citations	Purchased from Acris: Gruh I, Beilner J, Blomer U, Schmiedl A, Schmidt-Richter I, Kruse ML, et al. No evidence of transdifferentiation of human endothelial progenitor cells into cardiomyocytes after coculture with neonatal rat cardiomyocytes. Circulation. 2006 Mar 14;113(10):1326-34. Epub 2006 Mar 6. PubMed PMID: 16520414. Seibl R, Birchler T, Loeliger S, Hossle JP, Gay RE, Saurenmann T, et al. Expression and regulation of Toll-like receptor 2 in rheumatoid arthritis synovium. Am J Pathol. 2003 Apr;162(4):1221-7. PubMed PMID: 12651614. (Free PMC Article available, 4 images available) Zong W, Meyn LA, Moalli PA. The amount and activity of active matrix metalloproteinase 13 is suppressed by estradiol and progesterone in human pelvic floor fibroblasts. Biol Reprod. 2009 Feb;80(2):367-74. doi: 10.1095/biolreprod.108.072462. Epub 2008 Nov 5. PubMed PMID: 18987329. (Free PMC Article available, 6 images available) Beyer C, Schramm A, Akhmetshina A, Dees C, Kireva T, Gelse K, et al. β-catenin is a central mediator of pro-fibrotic Wnt signaling in systemic sclerosis. Ann Rheum Dis. 2012 May;71(5):761-7. doi: 10.1136/annrheumdis-2011-200568. Epub 2012 Feb 10. PubMed PMID: 22328737. (Free PMC Article available, 4 images available) Tomcik M, Palumbo-Zerr K, Zerr P, Sumova B, Avouac J, Dees C, et al. Tribbles homologue 3 stimulates canonical TGF-β signalling to regulate fibroblast activation and tissue fibrosis. Ann Rheum Dis. 2015 Jan 20. pii: annrheumdis-2014-206234. doi: 10.1136/annrheumdis-2014-206234. PubMed PMID: 25603829. Yamamura Y, Asai N, Enomoto A, Kato T, Mii S, Kondo Y, et al. Akt-Girdin signaling in cancer-associated fibroblasts contributes to tumor progression. Cancer Res. 2015 Mar 1;75(5):813-23. doi: 10.1158/0008-5472.CAN-14-1317. PubMed PMID: 25732845. Kodaira M, Hatakeyama H, Yuasa S, Seki T, Egashira T, Tohyama S, et al. Impaired respiratory function in MELAS-induced pluripotent stem cells with high heteroplasmy levels. FEBS Open Bio. 2015 Mar 20;5:219-25. doi: 10.1016/j.fob.2015.03.008. eCollection 2015. PubMed PMID: 25853038. (Free PMC Article available) Hashizume O, Ohnishi S, Mito T, Shimizu A, Iashikawa K, Nakada K, et al. Epigenetic regulation of the nuclear-coded GCAT and SHMT2 genes confers human age-associated mitochondrial respiration defects. Sci Rep. 2015 May 22;5:10434. doi: 10.1038/srep10434. PubMed PMID: 26000717.
General Readings	Bai Y, Muragaki Y, Obata K, Iwata K, Ooshima A. Immunological properties of monoclonal antibodies to human and rat prolyl 4-hydroxylase. J Biochem. 1986 Jun;99(6):1563-70. PubMed PMID: 3017922.
Storage	Prior to reconstitution store at 2-8°C. Following reconstitution store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch.
Format	Purification: Affinity Chromatography Buffer System: PBS, pH 7.2, containing 0.5% protease-free BSA, 0.09% Sodium Azide as preservative State: Lyophilized purified Ig fraction Purified Reconstitution: Restore in 0.5 ml double distilled water
Species Reactivity	Species reactivity (tested): Human
Specificity	Specificity: Clone 3-2B12 reacts with the beta subunit of Human P4H specifically found in fibroblasts. Has been reported to cross react with Rat P4H beta.

Accessory Products

Recommended Secondary Antibodies for Prolyl 4-hydroxylase subunit beta (11 products)

Catalog No.	Host	Pres.	Applications
R1253AP	Mouse IgG (H+L chain) antibody
	Rabbit	AP	C, E, WB
				OriGene Technologies GmbH
R1253B	Mouse IgG (H+L chain) antibody
	Rabbit	Biotin	C, E, WB
				OriGene Technologies GmbH
R1253F	Mouse IgG (H+L chain) antibody
	Rabbit	FITC	E, ICC/IF
				OriGene Technologies GmbH
R1253HRP	Mouse IgG (H+L chain) antibody
	Rabbit	HRP	C, E, WB
				OriGene Technologies GmbH
R1253TR	Mouse IgG (H+L chain) antibody
	Rabbit	Texas Red	ICC/IF
				OriGene Technologies GmbH
R1403R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
	Rabbit	PE	F
				OriGene Technologies GmbH
R1405R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
	Goat	PE
				OriGene Technologies GmbH
R1455R	Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody
	Donkey	PE
				OriGene Technologies GmbH
R1457C2	Mouse IgG (H+L chain), adsorbed antibody
	Goat	Cy2
				OriGene Technologies GmbH
R1457C3	Mouse IgG (H+L chain), adsorbed antibody
	Goat	Cy3
				OriGene Technologies GmbH
R1457C5	Mouse IgG (H+L chain), adsorbed antibody
	Goat	Cy5
				OriGene Technologies GmbH

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AF0910-1 Prolyl 4-hydroxylase subunit beta antibody

Quick Overview

Product Description for Prolyl 4-hydroxylase subunit beta

Properties for Prolyl 4-hydroxylase subunit beta

Datasheet Extract

Accessory Products

Recommended Secondary Antibodies for Prolyl 4-hydroxylase subunit beta (11 products)

Mouse IgG (H+L chain) antibody

Mouse IgG (H+L chain) antibody

Mouse IgG (H+L chain) antibody

Mouse IgG (H+L chain) antibody

Mouse IgG (H+L chain) antibody

Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody

Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody

Mouse IgG (H+L chain), F(ab)2 Fragment, adsorbed antibody

Mouse IgG (H+L chain), adsorbed antibody

Mouse IgG (H+L chain), adsorbed antibody

Mouse IgG (H+L chain), adsorbed antibody

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