Immunofluorescence analysis of mesenchymal stem cells using Zymed Ms anti-Stro-1 (Cat. No. 39-8401) and Gt anti-Mouse-Alexa Fluor 488 (Cat. No. A21042) (green). Tubulin is stained with phalloidin-Alexa 594 (red) and nuclei are stained with DAPI (blue). Sample is mounted in ProLong Gold antifade reagent.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Mouse Mu immunonglobin|
|Conjugate||Alexa Fluor® 488|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human IgGl, IgG2a, IgG2b, IgG3, IgA, human serum and purified human paraproteins prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-10 µg/mL|
|Immunofluorescence (IF)||1-10 µg/mL|
|Immunohistochemistry (IHC)||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-mouse IgM whole secondary antibodies have been affinity purified and cross-adsorbed against human IgGl, IgG2a, IgG2b, IgG3, IgA, human serum, and purified human paraproteins prior to conjugation. 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 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 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 488 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.
Inducible depletion of adult skeletal muscle stem cells impairs the regeneration of neuromuscular junctions.
A-21042 was used in immunohistochemistry to analyze impairment of the regeneration of neuromuscular junctions by an inducible depletion of adult skeletal muscle stem cells
|Liu W,Wei-LaPierre L,Klose A,Dirksen RT,Chakkalakal JV||eLife (4:null)||2015|
Mitochondrial morphology is altered in atrophied skeletal muscle of aged mice.
A-21042 was used in immunohistochemistry - frozen section to study atrophied skeletal muscle of aged mice and alteration of mitochondrial morphology
|Leduc-Gaudet JP,Picard M,St-Jean Pelletier F,Sgarioto N,Auger MJ,Vallée J,Robitaille R,St-Pierre DH,Gouspillou G||Oncotarget (6:17923)||2015|
|Not Applicable||Not Cited||
Immunization with malondialdehyde-modified low-density lipoprotein (LDL) reduces atherosclerosis in LDL receptor-deficient mice challenged with Porphyromonas gingivalis.
A-21042 was used in flow cytometry to study the effect of natural immune responses to malondialdehyde-modified LDL in protection against atherosclerosis.
|Turunen SP,Kummu O,Wang C,Harila K,Mattila R,Sahlman M,Pussinen PJ,Hörkkö S||Innate immunity (21:370)||2015|
|Not Applicable||Not Cited||Rescue of skeletal muscle alpha-actin-null mice by cardiac (fetal) alpha-actin.||Nowak KJ,Ravenscroft G,Jackaman C,Filipovska A,Davies SM,Lim EM,Squire SE,Potter AC,Baker E,Clément S,Sewry CA,Fabian V,Crawford K,Lessard JL,Griffiths LM,Papadimitriou JM,Shen Y,Morahan G,Bakker AJ,Davies KE,Laing NG||The Journal of cell biology (185:903)||2009|
|Not Applicable||Not Cited||Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos.||Lerou PH,Yabuuchi A,Huo H,Miller JD,Boyer LF,Schlaeger TM,Daley GQ||Nature protocols (3:923)||2008|
|Not Applicable||Not Cited||Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart.||Ott HC,Matthiesen TS,Goh SK,Black LD,Kren SM,Netoff TI,Taylor DA||Nature medicine (14:213)||2008|
|Not Applicable||Not Cited||Phospholipid biosynthesis program underlying membrane expansion during B-lymphocyte differentiation.||Fagone P,Sriburi R,Ward-Chapman C,Frank M,Wang J,Gunter C,Brewer JW,Jackowski S||The Journal of biological chemistry (282:7591)||2007|
|Not Applicable||Not Cited||Interaction of arginine-rich peptides with membrane-associated proteoglycans is crucial for induction of actin organization and macropinocytosis.||Nakase I,Tadokoro A,Kawabata N,Takeuchi T,Katoh H,Hiramoto K,Negishi M,Nomizu M,Sugiura Y,Futaki S||Biochemistry (46:492)||2007|
|Not Applicable||Not Cited||Cholera toxin B subunit binding does not correlate with GM1 expression: a study using mouse embryonic neural precursor cells.||Yanagisawa M,Ariga T,Yu RK||Glycobiology (16:19G)||2006|
|Not Applicable||Not Cited||Metabolic changes in glucose transporter-deficient Leishmania mexicana and parasite virulence.||Rodríguez-Contreras D,Landfear SM||The Journal of biological chemistry (281:20068)||2006|
|Not Applicable||Not Cited||Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons.||Murrey HE,Gama CI,Kalovidouris SA,Luo WI,Driggers EM,Porton B,Hsieh-Wilson LC||Proceedings of the National Academy of Sciences of the United States of America (103:21)||2006|
|Not Applicable||Not Cited||Four-color staining combining fluorescence and brightfield microscopy for simultaneous immune cell phenotyping and localization in tumor tissue sections.||van Vlierberghe RL,Sandel MH,Prins FA,van Iersel LB,van de Velde CJ,Tollenaar RA,Kuppen PJ||Microscopy research and technique (67:15)||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||Asymmetric localization of LGN but not AGS3, two homologs of Drosophila pins, in dividing human neural progenitor cells.||Fuja TJ,Schwartz PH,Darcy D,Bryant PJ||Journal of neuroscience research (75:782)||2004|
|Not Applicable||Not Cited||Muscle spindles and Golgi tendon organs in bovine calf extraocular muscle studied by means of double-fluorescent labeling, electron microscopy, and three-dimensional reconstruction.||Blumer R,Konakci KZ,Brugger PC,Blumer MJ,Moser D,Schoefer C,Lukas JR,Streicher J||Experimental eye research (77:447)||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||MyoD-positive myoblasts are present in mature fetal organs lacking skeletal muscle.||Gerhart J,Bast B,Neely C,Iem S,Amegbe P,Niewenhuis R,Miklasz S,Cheng PF,George-Weinstein M||The Journal of cell biology (155:381)||2001|