Immunofluorescent analysis of KDEL in U251 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a KDEL polyclonal antibody (Product # PA1-013) at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552). KDEL staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Published species reactivity||Rat, Hamster, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C T(643) G E E D T S E K D E L(654) of rat GRP94.|
|Purification||Antigen affinity chromatography|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-20 µg/ml|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||8-16 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-013 detects KDEL from human, rat, mouse and hamster.
PA1-013 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects a 57 kDa protein representing protein disulfide isomerase (PDI), a 78 kDa protein representing glucose regulated protein 70 kDa (GRP78) and an ~94 kDa protein representing GRP94 in mouse myeloma cells. Immunofluorescence staining of GRP78 in mouse myeloma cells with PA1-013 results in diffuse cytoplasmic staining consistent with specific ER localization. PA1-013 immunoprecipitates PDI, GRP78, an ~72 kDa protein which may be endoplasmic reticulum protein 72 kDa (ERp72), and an ~94 kDa protein representing GRP94 from numerous rodent tissues.
The PA1-013 immunogen is a synthetic peptide corresponding to residues C T(643) G E E D T S E K D E L(654) of rat GRP94.
Reconstitute with PBS.
The sequence Lys-Asp-Glu-Leu (KDEL) or a closely related sequence, is present at the carboxy-terminus of soluble endoplasmic reticulum (ER) resident proteins and some membrane proteins. 78 and 94 kDa glucose regulated proteins, GRP78 and GRP94 respectively, and protein disulfide isomerase (PDI) all share the C-terminal KDEL sequence. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Axons provide the secretory machinery for trafficking of voltage-gated sodium channels in peripheral nerve.
PA1-013 was used in immunocytochemistry to characterize the secretory machinery in axons and study its contribution to plasma membrane delivery of sodium channels
|González C,Cánovas J,Fresno J,Couve E,Court FA,Couve A||Proceedings of the National Academy of Sciences of the United States of America (113:1823)||2016|
Primary murine airway smooth muscle cells exposed to poly(I,C) or tunicamycin synthesize a leukocyte-adhesive hyaluronan matrix.
PA1-013 was used in western blot to study the response of primary murine airway smooth muscle cells to poly (I,C) or tunicamycin
|Lauer ME,Mukhopadhyay D,Fulop C,de la Motte CA,Majors AK,Hascall VC||The Journal of biological chemistry (284:5299)||2009|
Differentiated murine airway epithelial cells synthesize a leukocyte-adhesive hyaluronan matrix in response to endoplasmic reticulum stress.
PA1-013 was used in western blot to examine hyaluronan synthesis by the airway epithelial cells in response to endoplasmic reticulum stress.
|Lauer ME,Erzurum SC,Mukhopadhyay D,Vasanji A,Drazba J,Wang A,Fulop C,Hascall VC||The Journal of biological chemistry (283:26283)||2008|
Changes of endoplasmic reticulum chaperone complexes, redox state, and impaired protein disulfide reductase activity in misfolding alpha1-antitrypsin transgenic mice.
PA1-013 was used in western blot to investigate the changes of endoplasmic reticulum chaperone complexes, redox state, and impaired protein disulfide reductase activity in misfolding alpha1-antitrypsin transgenic mice
|Papp E,Száraz P,Korcsmáros T,Csermely P||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (20:1018)||2006|
Protein-disulfide isomerase-mediated reduction of the A subunit of cholera toxin in a human intestinal cell line.
PA1-013 was used in western blot to study the mechanisms of cholera toxin A subunit reduction in a human intestinal cell line
|Orlandi PA||The Journal of biological chemistry (272:4591)||1997|
Further characterization of mammalian ceramide kinase: substrate delivery and (stereo)specificity, tissue distribution, and subcellular localization studies.
PA1-013 was used in immunocytochemistry to investigate related factors affecting recombinant human ceramide kinase's activity
|Van Overloop H,Gijsbers S,Van Veldhoven PP||Journal of lipid research (47:268)||2006|
Functional proteomic screens reveal an essential extracellular role for hsp90 alpha in cancer cell invasiveness.
PA1-013 was used in immunoprecipitation to study the role of extracellular Hsp90-alpha in the activation of MMP2 and invasiveness of fibrosarcoma cells
|Eustace BK,Sakurai T,Stewart JK,Yimlamai D,Unger C,Zehetmeier C,Lain B,Torella C,Henning SW,Beste G,Scroggins BT,Neckers L,Ilag LL,Jay DG||Nature cell biology (6:507)||2004|