MAB1276 Anti-Nuclei & Chromosomes Antibody, histone H1 protein, clone 1415-1

MAB1276
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, R ELISA, WB, ICC, IH(P) M Purified Monoclonal Antibody
      Description
      Catalogue NumberMAB1276
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Nuclei & Chromosomes Antibody, histone H1 protein, clone 1415-1
      References
      Product Information
      FormatPurified
      Control
      • POSITIVE CONTROL:

        Human tonsil
      Presentation100 μL Hybridoma Supernatant, concentrated by ammonium sulfate. Buffer: PBS with 0.1% sodium azide.
      Applications
      ApplicationAnti-Nuclei & Chromosomes, histone H1 protein, clone 1415-1 is a high quality Mouse Monoclonal Antibody for the detection of Nuclei & Chromosomes & has been validated in ELISA, WB, ICC, IHC(P).
      Key Applications
      • ELISA
      • Western Blotting
      • Immunocytochemistry
      • Immunohistochemistry (Paraffin)
      Application NotesImmunocytochemistry on paraformaldehyde/acetone fixed cell preparation (indirect immunofluorescence) : 1:20-30, 30 - 100 μl per slide/well.

      Immunohistochemstry on formalin fixed, paraffin embedded tissue sections or paraformaldehyde/acetone fixed frozen tissue sections.

      Immunoblotting: non-reduced samples only, recognizes primarily 30-32kDa histone H1 band, other bands are present as well.

      ELISA: capture antibody when plated on to goat-anti-Ms Fc specific secondary, 1-4μg/mL



      Optimal working dilutions must be determined by end user.

      Subcellular Particle

      Suggested Protocol

      IMMUNOFLUORESCENCE AND ANTIBODY SCREENING PROCEDURE

      Hybridoma supernatants are examined by indirect immunofluorescence on cell preparations of human lymphnoid cells. In order to examine many samples in a short period of time, washed cells (wash 2 times in wash buffer at 4°C) at a concentration of 5 x 106 cells/mL in PBS are pipetted dropwise on PTFE coated printed microscope slides containing ten 5 mm wells/slide. After the cells are allowed to settle to the surface of the glass (10-15 minutes only), the overlying fluid is quickly removed by aspiration and the cells are dried to the slide by a gentle stream of warm air. The slides are then immediately fixed in 2% formaldehyde, ultra-pure, in PBS for 15 minutes at room temperature. After fixation, the slides are rinsed in PBS and placed in acetone at -20°C for 3 minutes to make the cells permeable. After a final rinse in PBS to remove the acetone, the slides are stored in PBS at 4°C indefinitely in covered Coplan jars.

      In addition to the lymphnoid cultures, normal human epithelial cells can be screened by indirect immunofluorescence microscopy for positive reactions with the hybridoma supernatants. Since the human epithelial cells grow as monolayer cultures, they are plated directly onto the printed microscope slides after trypsinization and allowed to attach and grow overnight at 37°C in complete medium. The following day, the slides are briefly rinsed in PBS to remove the medium and the cells are fixed as described above. In general, the slides are not allowed to air dry either during or after the fixation procedure in order to maintain the cellular integrity and antigenicity of intracellular molecules.

      For photographic analysis, viable cell preparations obtained from Ficoll™-hypaque gradient separations are cytocentrifuged directly onto slides at 1250 rpm for 10 minutes. This procedure flattens the lymphnoid cells and greatly improves the visibility of intranuclear and cytoplasmic antigens. Slides prepared in this manner are fixed in the same way directly after cytocentrifugation.

      In order to screen the hybridoma supernatants by indirect immunofluorescence, 30-100 μL of each supernatant (optimize for each individual assay) are pipetted on a well of the printed microscope slides using a different tip for each supernatant. After 60 minutes of incubation at 37°C in a humidified chamber, the slides are rinsed 3 times with PBS at room temperature, and again incubated for 30 minutes at 37°C with 20 μL of a 1:20 dilution of fluorescein-conjugated goat anti-mouse IgG antibody (Millipore AP124F). The slides are then prepared rinsed 3 times with PBS, counterstained with Evans Blue for 5 minutes at room temperature using a freshly prepared solution containing 50 μL of a 1% stock solution of Evans Blue in 80 mL of PBS, rinsed a final time in PBS, and coverslipped using a 1:1 solution of glycerol: PBS. The slides are then examined by epifluorescence microscopy. Since many of the monoclonal antibodies produced a rapidly diminishing fluorescent reaction, exposure times optimally are less than five seconds.

      Important Note: During shipment, small volumes of antibody will occasionally become entrapped in the seal of the product vial. For antibodies with volumes of 200 μl or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container's cap.
      Biological Information
      Epitopehistone H1 protein
      Clone1415-1
      HostMouse
      SpecificityHistone specific; stains nuclei and chromosomes of cells of all species including animals and plants. Reacts with preferentially with Histone H1 by ELISA and immunoblotting. MAB1276 produces a speckled nuclear staining pattern in all tissues studies.

      Stains nuclei of all human cell types and also stains chromosomes diffusely in metaphase cells. Nuclear staining in interphase cells is intense and diffuse. Antibody reacts with rat cells {Bendeck, 2001}.
      IsotypeIgG2a
      Species Reactivity
      • Human
      • Rat
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins responsible for nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a member of the histone H1 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6.
      Gene Symbol
      • HIST1H1E
      • H1.4
      • dJ221C16.5
      • H1F4
      • MGC116819
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P10412 # Histones H1 are necessary for the condensation of nucleosome chains into higher order structures.
      SIZE: 219 amino acids; 21865 Da
      SUBCELLULAR LOCATION: Nucleus.
      SIMILARITY: SwissProt: P10412 ## Belongs to the histone H1/H5 family.
      MISCELLANEOUS: This variant accounts for 60% of histone H1.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain at -20°C for up to 12 months. Avoid repeated freeze/thaw cycles.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-Nuclei & Chromosomes Antibody, histone H1 protein, clone 1415-1 SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-Nuclei & Chromosomes Antibody, histone H1 protein, clone 1415-1 Certificates of Analysis

      TitleLot Number
      Anti-Nuclei Chromosomes, histone H1 protein, clone 1415-1 - 19973581997358
      Anti-Nuclei Chromosomes, histone H1 protein, clone 1415-1 - NG1869832NG1869832
      Anti-Nuclei Chromosomes, histone H1 protein, clone 1415-1 - NG1899913NG1899913
      Anti-Nuclei & Chromosomes, histone -25521182552118
      Anti-Nuclei & Chromosomes, histone -Q2630368Q2630368
      Anti-Nuclei & Chromosomes, histone H1 protein, clone 1415-1 - 25584262558426
      Anti-Nuclei & Chromosomes, histone H1 protein, clone 1415-1 Monoclonal Antibody3123247
      Anti-Nuclei & Chromosomes, histone H1 protein, clone 1415-1 Monoclonal Antibody3130528
      Anti-Nuclei & Chromosomes, histone H1 protein, clone 1415-1 Monoclonal Antibody3084132

      References

      Reference overviewPub Med ID
      15-Lipoxygenase-1-enhanced Src-Janus kinase 2-signal transducer and activator of transcription 3 stimulation and monocyte chemoattractant protein-1 expression require redox-sensitive activation of epidermal growth factor receptor in vascular wall remodeling.
      Singh, NK; Wang, D; Kundumani-Sridharan, V; Van Quyen, D; Niu, J; Rao, GN
      The Journal of biological chemistry  286  22478-88  2011

      Show Abstract
      21536676 21536676
      Insulin increases reendothelialization and inhibits cell migration and neointimal growth after arterial injury.
      Danna M Breen, Kalam K Chan, Jiwanjeet K Dhaliwall, Michael R Ward, Nael Al Koudsi, Loretta Lam, Melissa De Souza, Husam Ghanim, Paresh Dandona, Duncan J Stewart, Michelle P Bendeck, Adria Giacca
      Arteriosclerosis, thrombosis, and vascular biology  29  1060-6  2009

      Show Abstract
      19359661 19359661
      A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction.
      Pasierbek, P; Jantsch, M; Melcher, M; Schleiffer, A; Schweizer, D; Loidl, J
      Genes & development  15  1349-60  2001

      Show Abstract Full Text Article
      11390355 11390355
      Preparation and imaging of nuclear spreads from cells of the zebrafish embryo. Evidence for large degradation intermediates in apoptosis.
      D W Chan, T D Yager
      Chromosoma  107  39-60  1998

      Show Abstract
      9567200 9567200
      The Drosophila suppressor of sable protein binds to RNA and associates with a subset of polytene chromosome bands.
      M V Murray, M A Turnage, K J Williamson, W R Steinhauer, L L Searles, M V Murray, M A Turnage, K J Williamson, W R Steinhauer, L L Searles
      Molecular and cellular biology  17  2291-300  1997

      Show Abstract Full Text Article
      9121479 9121479
      High-resolution imaging at the cellular and subcellular levels in flattened whole mounts of early zebrafish embryos.
      T D Yager, R Ikegami, A K Rivera-Bennetts, C Zhao, D Brooker
      Biochemistry and cell biology = Biochimie et biologie cellulaire  75  535-50  1997

      Show Abstract
      9551178 9551178
      Activation of the metaphase checkpoint and an apoptosis programme in the early zebrafish embryo, by treatment with the spindle-destabilising agent nocodazole.
      R Ikegami, J Zhang, A K Rivera-Bennetts, T D Yager
      Zygote (Cambridge, England)  5  329-50  1997

      Show Abstract
      9563681 9563681

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      Life Science Research > Antibodies and Assays > Primary Antibodies