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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R | ELISA, WB, ICC, IH(P) | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB1276 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-Nuclei & Chromosomes Antibody, histone H1 protein, clone 1415-1 |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | 100 μL Hybridoma Supernatant, concentrated by ammonium sulfate. Buffer: PBS with 0.1% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-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 |
|
| Application Notes | Immunocytochemistry 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 containers cap. |
| Biological Information | |
|---|---|
| Epitope | histone H1 protein |
| Clone | 1415-1 |
| Host | Mouse |
| Specificity | Histone 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}. |
| Isotype | IgG2a |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | Histones 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 |
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| UniProt Number |
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| UniProt Summary | FUNCTION: 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. |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at -20°C for up to 12 months. Avoid repeated freeze/thaw cycles. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |