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Product Description for Prolyl-4-Hydroxylase beta
Properties for Prolyl-4-Hydroxylase beta
| Manufacturer | Acris Antibodies GmbH |
| Quantity | 0.1 mg |
| Reactivity | Human |
| Presentation | Purified |
| Applications | Frozen sections, Paraffin Sections, Western blot / Immunoblot |
| Isotype | IgG1 |
| Host | Mouse |
| Clone | 3-2B12 |
| Shipping to | Worldwide |
Datasheet Extract
| Synonyms | Fibroblast Marker, EC=5.3.4.1, Protein Disulfide-Isomerase, p55, P4HB, ERBA2L, PDI, PDIA1, PO4DB, FocusOn008 |
| Immunogen | Native human merosin M-chain |
| Product Type | Antibodies |
| Recommended Isotype Control | SM10P (for use in human samples), AM03095PU-N |
| Immunogen | Remarks: The epitope has not been further determined. |
| Storage | Original vial: One year from despatch at 2-8°C. Stock solution or aliquots thereof: One year at -20°C. Avoid repeated thawing and freezing. |
| References | 1. Bai, Y. et al.: Immunological properties of monoclonal antibodies to human and rat prolyl 4-hydroxylase. J. Biochem. 99, 1563-70 (1986). |
| Format | Reconstitution: Restore with 0.5 ml distilled water. |
| State: Lyophilized purified Ig fraction | |
| Purification: Affinity Chromatography | |
| Buffer System: PBS, pH 7.2, containing 5 mg/ml BSA as a stabilizer and 0.09% Sodium Azide as preservative. | |
| Protocols | Protocol for Frozen Sections: Incubations are done at room temperature. Water is of double distilled or comparable quality. Fix fresh frozen sections in ice-cold acetone for 10 min Block endogenous peroxidase with 100ml 0.15M sodium azide / 0.15% H2O2 in PBS Wash in PBS Block with 10% Normal Rabbit Serum for 30 min in a humid chamber Incubate with primary antibody #AF0910-1 for 1 hour in a humid chamber Wash in PBS Incubate with secondary antibody (peroxidase-conjugated Rabbit Anti Mouse IgG Cat# R1253HRP), at the recommended dilution for 1 hour in a humid chamber Wash in PBS Incubate with AEC substrate (3-Amino-9-Ethylcarbazol) for 12 minutes Wash in PBS Counterstain with Mayer's hemalum Protocol for Paraffin Sections: Incubations are done at room temperature. Water is of double distilled or comparable quality. Rehydrate paraffin sections Put the slides in a cuvette with 250 ml 0.01 M citrate buffer pH 6.0 Heat the slides in a microwave oven for 2 x 7 min and 700 Watt Leave the slides in the buffer for 20 min Block endogenous peroxidase with 1% H2O2 in water Wash in PBS Block with 10% Normal Goat Serum for 30 min in a humid chamber Incubate with primary antibody #AF0910-1 for 1 hour in a humid chamber Wash in PBS 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 Incubate with AEC substrate (3-Amino-9-Ethylcarbazol) for 12 minutes Wash in PBS Counterstain |
| Specificity | Clone 3-2B12 reacts with the beta subunit of human P4H specifically found in fibroblasts. |
| Add. information | Has been reported to cross react with rat P4H beta in Western blot 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) |
| Species Reactivity | Tested: Human |
| Focuson | 008 Fibroblast Marker Antibodies - FocusOn No.8 |
Secondary Antibodies for Prolyl-4-Hydroxylase beta
| Catalog No. | Name / Host | Presentation | Reactivity | ||||
|---|---|---|---|---|---|---|---|
| R1253AP | Mouse IgG (H&L) | ||||||
| Rabbit | AP | 1 mg / 190 Euro | |||||
| R1253B | Mouse IgG (H&L) | ||||||
| Rabbit | Biotin | 2 mg / 170 Euro | |||||
| R1253F | Mouse IgG (H&L) | ||||||
| Rabbit | FITC | 2 mg / 160 Euro | |||||
| R1253HRP | Mouse IgG (H&L) | ||||||
| Rabbit | HRP | 2 mg / 180 Euro | |||||
| R1253TR | Mouse IgG (H&L) | ||||||
| Rabbit | Texas Red | 2 mg / 170 Euro | |||||
| R1403R | Mouse IgG (H&L) F(ab')2 Fragment | ||||||
| Rabbit | PE | 1 ml / 290 Euro | |||||
| R1405R | Mouse IgG (H&L) F(ab')2 Fragment | ||||||
| Goat | PE | 1 ml / 290 Euro | |||||
| R1455R | Mouse IgG (H&L) F(ab')2 Fragment multi-species ads. | ||||||
| Donkey | PE | 1 ml / 290 Euro | |||||
| R1457C2 | Mouse IgG (H&L) multi-species ads. | ||||||
| Goat | Cy2 | 1 mg / 270 Euro | |||||
| R1457C3 | Mouse IgG (H&L) multi-species ads. | ||||||
| Goat | Cy3 | 1 mg / 250 Euro | |||||
| R1457C5 | Mouse IgG (H&L) multi-species ads. | ||||||
| Goat | Cy5 | 1 mg / 250 Euro | |||||
