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Immunofluorescent analysis of Phospho-RAD17 pSer646 showing staining in the nucleus of HCT116 cells. HCT116 cells were fixed in 4% paraformaldehyde at RT for 15 min and stained using a Phospho-RAD17 pSer646 polyclonal antibody (Product # PA5-34970) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 546|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against bovine IgG, goat IgG, mouse IgG, rat IgG and human IgG|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunofluorescence (IF)||1-10 µg/mL|
|Immunohistochemistry (Frozen) (IHC (F))||1-10 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.
To minimize cross-reactivity, these goat anti-rabbit IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against bovine IgG, goat IgG, mouse IgG, rat IgG, and human IgG. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.
Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 546 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 546 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 546 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 546 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection. The degree of labeling for each conjugate is typically 2-8 fluorophore molecules per IgG molecule; the exact degree of labeling is indicated on the certificate of analysis for each product lot.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically. For the fluorophore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.
We offer an extensive line of Invitrogen™ secondary antibody conjugates with well-characterized specificity and labeled with a wide selection of premium fluorescent dyes, including Invitrogen™ Alexa Fluor™ fluorescent dyes. Fluorescent secondary antibody conjugates are useful in the detection, sorting, or purification of its specified target and ideal for fluorescence microscopy and confocal laser scanning microscopy, flow cytometry, and fluorescent western detection. The breadth of fluorescent markers we offer allows our reagents to be tailored to almost any fluorescent detection system.
Secondary antibodies may be provided in three formats: whole IgG, divalent F(ab')2 fragments, and monovalent Fab fragments. Because of the high degree of conservation in the structure of many immunoglobulin domains, most class-specific secondary antibodies must be affinity-purified and cross-adsorbed to achieve minimal cross-reaction with other immunoglobulins.
Our secondary antibody conjugates are most commonly prepared by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (e.g., immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. In the first round of purification, whole immunoglobulins binding to the immunizing antibody are recovered and mainly consist of the ~150-kDa IgG class. Further purification, for example, with Protein A or G, removes all unwanted immunoglobulin classes except the affinity-purified antibodies that react with the target-specific immunoglobulin heavy and/or light chains.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Gene therapy restores vision in rd1 mice after removal of a confounding mutation in Gpr179.
A-11035 was used in immunohistochemistry - frozen section to investigate how the Grp179 mutation contributes to bipolar cell-mediated b-waves in mice.
|Nishiguchi KM,Carvalho LS,Rizzi M,Powell K,Holthaus SM,Azam SA,Duran Y,Ribeiro J,Luhmann UF,Bainbridge JW,Smith AJ,Ali RR||Nature communications (6:null)||2015|
|Not Applicable||Not Cited||Amino acid residues critical for endoplasmic reticulum export and trafficking of platelet-activating factor receptor.||Hirota N,Yasuda D,Hashidate T,Yamamoto T,Yamaguchi S,Nagamune T,Nagase T,Shimizu T,Nakamura M||The Journal of biological chemistry (285:5931)||2010|
|Not Applicable||Not Cited||Visualization and identification of IL-7 producing cells in reporter mice.||Mazzucchelli RI,Warming S,Lawrence SM,Ishii M,Abshari M,Washington AV,Feigenbaum L,Warner AC,Sims DJ,Li WQ,Hixon JA,Gray DH,Rich BE,Morrow M,Anver MR,Cherry J,Naf D,Sternberg LR,McVicar DW,Farr AG,Germain RN,Rogers K,Jenkins NA,Copeland NG,Durum SK||PloS one (4:null)||2009|
|Not Applicable||Not Cited||Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro.||Mezaki Y,Yamaguchi N,Yoshikawa K,Miura M,Imai K,Itoh H,Senoo H||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (57:687)||2009|
|Not Applicable||Not Cited||Chitin is a size-dependent regulator of macrophage TNF and IL-10 production.||Da Silva CA,Chalouni C,Williams A,Hartl D,Lee CG,Elias JA||Journal of immunology (Baltimore, Md. : 1950) (182:3573)||2009|
|Not Applicable||Not Cited||pLG72 modulates intracellular D-serine levels through its interaction with D-amino acid oxidase: effect on schizophrenia susceptibility.||Sacchi S,Bernasconi M,Martineau M,Mothet JP,Ruzzene M,Pilone MS,Pollegioni L,Molla G||The Journal of biological chemistry (283:22244)||2008|
|Not Applicable||Not Cited||TRPC channels determine human keratinocyte differentiation: new insight into basal cell carcinoma.||Beck B,Lehen'kyi V,Roudbaraki M,Flourakis M,Charveron M,Bordat P,Polakowska R,Prevarskaya N,Skryma R||Cell calcium (43:492)||2008|
|Not Applicable||Not Cited||Impaired insulin secretion and glucose intolerance in synaptotagmin-7 null mutant mice.||Gustavsson N,Lao Y,Maximov A,Chuang JC,Kostromina E,Repa JJ,Li C,Radda GK,Südhof TC,Han W||Proceedings of the National Academy of Sciences of the United States of America (105:3992)||2008|
|Not Applicable||Not Cited||Selective photoinactivation of protein function through environment-sensitive switching of singlet oxygen generation by photosensitizer.||Yogo T,Urano Y,Mizushima A,Sunahara H,Inoue T,Hirose K,Iino M,Kikuchi K,Nagano T||Proceedings of the National Academy of Sciences of the United States of America (105:28)||2008|
|Not Applicable||Not Cited||BHK cells transfected with NCX3 are more resistant to hypoxia followed by reoxygenation than those transfected with NCX1 and NCX2: Possible relationship with mitochondrial membrane potential.||Secondo A,Staiano RI,Scorziello A,Sirabella R,Boscia F,Adornetto A,Valsecchi V,Molinaro P,Canzoniero LM,Di Renzo G,Annunziato L||Cell calcium (42:521)||2007|
|Not Applicable||Not Cited||Cholesterol controls lipid endocytosis through Rab11.||Takahashi M,Murate M,Fukuda M,Sato SB,Ohta A,Kobayashi T||Molecular biology of the cell (18:2667)||2007|
|Not Applicable||Not Cited||Specificity of olfactory receptor interactions with other G protein-coupled receptors.||Bush CF,Jones SV,Lyle AN,Minneman KP,Ressler KJ,Hall RA||The Journal of biological chemistry (282:19042)||2007|
|Not Applicable||Not Cited||Direct visualization of glucocorticoid receptor positive cells in the hippocampal regions using green fluorescent protein transgenic mice.||Nishi M,Usuku T,Itose M,Fujikawa K,Hosokawa K,Matsuda KI,Kawata M||Neuroscience (146:1555)||2007|
|Not Applicable||Not Cited||Post-translational arginylation of calreticulin: a new isospecies of calreticulin component of stress granules.||Decca MB,Carpio MA,Bosc C,Galiano MR,Job D,Andrieux A,Hallak ME||The Journal of biological chemistry (282:8237)||2007|
|Not Applicable||Not Cited||Neurotrophin 3 induces structural and functional modification of synapses through distinct molecular mechanisms.||Je HS,Yang F,Zhou J,Lu B||The Journal of cell biology (175:1029)||2006|
|Not Applicable||Not Cited||The P2Y12 receptor regulates microglial activation by extracellular nucleotides.||Haynes SE,Hollopeter G,Yang G,Kurpius D,Dailey ME,Gan WB,Julius D||Nature neuroscience (9:1512)||2006|
|Not Applicable||Not Cited||Cell-surface heparan sulfate proteoglycans are essential components of the unconventional export machinery of FGF-2.||Zehe C,Engling A,Wegehingel S,Schäfer T,Nickel W||Proceedings of the National Academy of Sciences of the United States of America (103:15479)||2006|
|Not Applicable||Not Cited||LMP4 regulates Tbx5 protein subcellular localization and activity.||Camarata T,Bimber B,Kulisz A,Chew TL,Yeung J,Simon HG||The Journal of cell biology (174:339)||2006|
|Not Applicable||Not Cited||Purification, characterization, and cloning of a Spodoptera frugiperda Sf9 beta-N-acetylhexosaminidase that hydrolyzes terminal N-acetylglucosamine on the N-glycan core.||Tomiya N,Narang S,Park J,Abdul-Rahman B,Choi O,Singh S,Hiratake J,Sakata K,Betenbaugh MJ,Palter KB,Lee YC||The Journal of biological chemistry (281:19545)||2006|
|Not Applicable||Not Cited||Expression of a functional Drosophila melanogaster CMP-sialic acid synthetase. Differential localization of the Drosophila and human enzymes.||Viswanathan K,Tomiya N,Park J,Singh S,Lee YC,Palter K,Betenbaugh MJ||The Journal of biological chemistry (281:15929)||2006|
|Not Applicable||Not Cited||Subcellular targeting of RGS9-2 is controlled by multiple molecular determinants on its membrane anchor, R7BP.||Song JH,Waataja JJ,Martemyanov KA||The Journal of biological chemistry (281:15361)||2006|
|Not Applicable||Not Cited||Systematic characterization of nuclear proteome during apoptosis: a quantitative proteomic study by differential extraction and stable isotope labeling.||Hwang SI,Lundgren DH,Mayya V,Rezaul K,Cowan AE,Eng JK,Han DK||Molecular & cellular proteomics : MCP (5:1131)||2006|
|Not Applicable||Not Cited||Glycogen synthase kinase-3beta inhibits the xenobiotic and antioxidant cell response by direct phosphorylation and nuclear exclusion of the transcription factor Nrf2.||Salazar M,Rojo AI,Velasco D,de Sagarra RM,Cuadrado A||The Journal of biological chemistry (281:14841)||2006|
|Not Applicable||Not Cited||The phosphorylation of serine 492 of perilipin a directs lipid droplet fragmentation and dispersion.||Marcinkiewicz A,Gauthier D,Garcia A,Brasaemle DL||The Journal of biological chemistry (281:11901)||2006|
|Not Applicable||Not Cited||The adaptor protein Nck interacts with Fas ligand: Guiding the death factor to the cytotoxic immunological synapse.||Lettau M,Qian J,Linkermann A,Latreille M,Larose L,Kabelitz D,Janssen O||Proceedings of the National Academy of Sciences of the United States of America (103:5911)||2006|
|Not Applicable||Not Cited||Intercellular spreading of Porphyromonas gingivalis infection in primary gingival epithelial cells.||Yilmaz O,Verbeke P,Lamont RJ,Ojcius DM||Infection and immunity (74:703)||2006|
|Not Applicable||Not Cited||Distinct regulation of Ubc13 functions by the two ubiquitin-conjugating enzyme variants Mms2 and Uev1A.||Andersen PL,Zhou H,Pastushok L,Moraes T,McKenna S,Ziola B,Ellison MJ,Dixit VM,Xiao W||The Journal of cell biology (170:745)||2005|
|Not Applicable||Not Cited||Connexin 43 interacts with zona occludens-1 and -2 proteins in a cell cycle stage-specific manner.||Singh D,Solan JL,Taffet SM,Javier R,Lampe PD||The Journal of biological chemistry (280:30416)||2005|
|Not Applicable||Not Cited||In vivo characterization of bone marrow-derived fibroblasts recruited into fibrotic lesions.||Ishii G,Sangai T,Sugiyama K,Ito T,Hasebe T,Endoh Y,Magae J,Ochiai A||Stem cells (Dayton, Ohio) (23:699)||2005|
|Not Applicable||Not Cited||Canonical transient receptor potential 1 plays a role in basic fibroblast growth factor (bFGF)/FGF receptor-1-induced Ca2+ entry and embryonic rat neural stem cell proliferation.||Fiorio Pla A,Maric D,Brazer SC,Giacobini P,Liu X,Chang YH,Ambudkar IS,Barker JL||The Journal of neuroscience : the official journal of the Society for Neuroscience (25:2687)||2005|
|Not Applicable||Not Cited||Improvement in nuclear entry and transgene expression of baculoviruses by disintegration of microtubules in human hepatocytes.||Salminen M,Airenne KJ,Rinnankoski R,Reimari J,Välilehto O,Rinne J,Suikkanen S,Kukkonen S,Ylä-Herttuala S,Kulomaa MS,Vihinen-Ranta M||Journal of virology (79:2720)||2005|
|Not Applicable||Not Cited||Receptor tyrosine phosphatase-dependent cytoskeletal remodeling by the hedgehog-responsive gene MIM/BEG4.||Gonzalez-Quevedo R,Shoffer M,Horng L,Oro AE||The Journal of cell biology (168:453)||2005|
|Not Applicable||Not Cited||Vaccinia virus F1L protein is a tail-anchored protein that functions at the mitochondria to inhibit apoptosis.||Stewart TL,Wasilenko ST,Barry M||Journal of virology (79:1084)||2005|
|Not Applicable||Not Cited||Precise localization of alpha7 nicotinic acetylcholine receptors on glutamatergic axon terminals in the rat ventral tegmental area.||Jones IW,Wonnacott S||The Journal of neuroscience : the official journal of the Society for Neuroscience (24:11244)||2004|
|Not Applicable||Not Cited||Protein kinase Calpha activates c-Src and induces podosome formation via AFAP-110.||Gatesman A,Walker VG,Baisden JM,Weed SA,Flynn DC||Molecular and cellular biology (24:7578)||2004|
|Not Applicable||Not Cited||Differential localization of the centromere-specific proteins in the major centromeric satellite of Arabidopsis thaliana.||Shibata F,Murata M||Journal of cell science (117:2963)||2004|
|Not Applicable||Not Cited||Stromal cell-derived factor-1alpha promotes melanoma cell invasion across basement membranes involving stimulation of membrane-type 1 matrix metalloproteinase and Rho GTPase activities.||Bartolomé RA,Gálvez BG,Longo N,Baleux F,Van Muijen GN,Sánchez-Mateos P,Arroyo AG,Teixidó J||Cancer research (64:2534)||2004|
|Not Applicable||Not Cited||Compaction of the Escherichia coli nucleoid caused by Cyt1Aa.||Manasherob R,Zaritsky A,Metzler Y,Ben-Dov E,Itsko M,Fishov I||Microbiology (Reading, England) (149:3553)||2003|
|Not Applicable||Not Cited||The B cell-specific major raft protein, Raftlin, is necessary for the integrity of lipid raft and BCR signal transduction.||Saeki K,Miura Y,Aki D,Kurosaki T,Yoshimura A||The EMBO journal (22:3015)||2003|
|Not Applicable||Not Cited||Phosphorylation of histone H2AX and activation of Mre11, Rad50, and Nbs1 in response to replication-dependent DNA double-strand breaks induced by mammalian DNA topoisomerase I cleavage complexes.||Furuta T,Takemura H,Liao ZY,Aune GJ,Redon C,Sedelnikova OA,Pilch DR,Rogakou EP,Celeste A,Chen HT,Nussenzweig A,Aladjem MI,Bonner WM,Pommier Y||The Journal of biological chemistry (278:20303)||2003|
|Not Applicable||Not Cited||Casein kinase I epsilon enhances the binding of Dvl-1 to Frat-1 and is essential for Wnt-3a-induced accumulation of beta-catenin.||Hino S,Michiue T,Asashima M,Kikuchi A||The Journal of biological chemistry (278:14066)||2003|
|Not Applicable||Not Cited||Expression of hornerin in stratified squamous epithelium in the mouse: a comparative analysis with profilaggrin.||Makino T,Takaishi M,Toyoda M,Morohashi M,Huh NH||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (51:485)||2003|
|Not Applicable||Not Cited||MTA1 interacts with MAT1, a cyclin-dependent kinase-activating kinase complex ring finger factor, and regulates estrogen receptor transactivation functions.||Talukder AH,Mishra SK,Mandal M,Balasenthil S,Mehta S,Sahin AA,Barnes CJ,Kumar R||The Journal of biological chemistry (278:11676)||2003|
|Not Applicable||Not Cited||Amassin, an olfactomedin protein, mediates the massive intercellular adhesion of sea urchin coelomocytes.||Hillier BJ,Vacquier VD||The Journal of cell biology (160:597)||2003|
|Not Applicable||Not Cited||Chlorotoxin inhibits glioma cell invasion via matrix metalloproteinase-2.||Deshane J,Garner CC,Sontheimer H||The Journal of biological chemistry (278:4135)||2003|
|Not Applicable||Not Cited||Localization of a mitochondrial type of NADP-dependent isocitrate dehydrogenase in kidney and heart of rat: an immunocytochemical and biochemical study.||Haraguchi CM,Mabuchi T,Yokota S||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (51:215)||2003|
|Not Applicable||Not Cited||Survivin enhances Aurora-B kinase activity and localizes Aurora-B in human cells.||Chen J,Jin S,Tahir SK,Zhang H,Liu X,Sarthy AV,McGonigal TP,Liu Z,Rosenberg SH,Ng SC||The Journal of biological chemistry (278:486)||2003|
|Not Applicable||Not Cited||Specific recruitment of human cohesin to laser-induced DNA damage.||Kim JS,Krasieva TB,LaMorte V,Taylor AM,Yokomori K||The Journal of biological chemistry (277:45149)||2002|
|Not Applicable||Not Cited||An ATP-binding cassette transporter is a major glycoprotein of sea urchin sperm membranes.||Mengerink KJ,Vacquier VD||The Journal of biological chemistry (277:40729)||2002|
|Not Applicable||Not Cited||SHP-2 tyrosine phosphatase as an intracellular target of Helicobacter pylori CagA protein.||Higashi H,Tsutsumi R,Muto S,Sugiyama T,Azuma T,Asaka M,Hatakeyama M||Science (New York, N.Y.) (295:683)||2002|
|Not Applicable||Not Cited||Small cargo proteins and large aggregates can traverse the Golgi by a common mechanism without leaving the lumen of cisternae.||Mironov AA,Beznoussenko GV,Nicoziani P,Martella O,Trucco A,Kweon HS,Di Giandomenico D,Polishchuk RS,Fusella A,Lupetti P,Berger EG,Geerts WJ,Koster AJ,Burger KN,Luini A||The Journal of cell biology (155:1225)||2001|
|Not Applicable||Not Cited||Role of heparan sulfate as a tissue-specific regulator of FGF-4 and FGF receptor recognition.||Allen BL,Filla MS,Rapraeger AC||The Journal of cell biology (155:845)||2001|
|Not Applicable||Not Cited||Directed polar secretion of protease from single cells of Vibrio cholerae via the type II secretion pathway.||Scott ME,Dossani ZY,Sandkvist M||Proceedings of the National Academy of Sciences of the United States of America (98:13978)||2001|
|Not Applicable||Not Cited||Distinct retrieval and retention mechanisms are required for the quality control of endoplasmic reticulum protein folding.||Vashist S,Kim W,Belden WJ,Spear ED,Barlowe C,Ng DT||The Journal of cell biology (155:355)||2001|
|Not Applicable||Not Cited||MKP-7, a novel mitogen-activated protein kinase phosphatase, functions as a shuttle protein.||Masuda K,Shima H,Watanabe M,Kikuchi K||The Journal of biological chemistry (276:39002)||2001|
|Not Applicable||Not Cited||Synergistic activation of the Wnt signaling pathway by Dvl and casein kinase Iepsilon.||Kishida M,Michiue T,Yamamoto H,Kishida S,Fukui A,Asashima M,Kikuchi A||The Journal of biological chemistry (276:33147)||2001|
|Not Applicable||Not Cited||Extranuclear lipid bodies, elicited by CCR3-mediated signaling pathways, are the sites of chemokine-enhanced leukotriene C4 production in eosinophils and basophils.||Bandeira-Melo C,Phoofolo M,Weller PF||The Journal of biological chemistry (276:22779)||2001|
|Not Applicable||Not Cited||A two-receptor pathway for catabolism of Clara cell secretory protein in the kidney.||Burmeister R,Boe IM,Nykjaer A,Jacobsen C,Moestrup SK,Verroust P,Christensen EI,Lund J,Willnow TE||The Journal of biological chemistry (276:13295)||2001|
|Not Applicable||Not Cited||Anaphase-promoting complex/cyclosome-dependent proteolysis of human cyclin A starts at the beginning of mitosis and is not subject to the spindle assembly checkpoint.||Geley S,Kramer E,Gieffers C,Gannon J,Peters JM,Hunt T||The Journal of cell biology (153:137)||2001|
|Not Applicable||Not Cited||Processing of beta-secretase by furin and other members of the proprotein convertase family.||Creemers JW,Ines Dominguez D,Plets E,Serneels L,Taylor NA,Multhaup G,Craessaerts K,Annaert W,De Strooper B||The Journal of biological chemistry (276:4211)||2001|
|Not Applicable||Not Cited||The platelet integrin alpha IIbbeta 3 has an endogenous thiol isomerase activity.||O'Neill S,Robinson A,Deering A,Ryan M,Fitzgerald DJ,Moran N||The Journal of biological chemistry (275:36984)||2000|
|Not Applicable||Not Cited||Properties of gap junction channels formed by Cx46 alone and in combination with Cx50.||Hopperstad MG,Srinivas M,Spray DC||Biophysical journal (79:1954)||2000|
|Not Applicable||Not Cited||The co-repressor mSin3A is a functional component of the REST-CoREST repressor complex.||Grimes JA,Nielsen SJ,Battaglioli E,Miska EA,Speh JC,Berry DL,Atouf F,Holdener BC,Mandel G,Kouzarides T||The Journal of biological chemistry (275:9461)||2000|
|Not Applicable||Not Cited||Novel functions of nanos in downregulating mitosis and transcription during the development of the Drosophila germline.||Deshpande G,Calhoun G,Yanowitz JL,Schedl PD||Cell (99:271)||1999|
|Not Applicable||Not Cited||Human bleomycin hydrolase binds ribosomal proteins.||Koldamova RP,Lefterov IM,DiSabella MT,Almonte C,Watkins SC,Lazo JS||Biochemistry (38:7111)||1999|