Human NMNAT-1 Antibody (675112)

R&D Systems | Catalog # MAB5865

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry

Cited:

Immunohistochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 675112
View all NMNAT-1 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human NMNAT-1
Met1-Thr279
Accession # Q9HAN9

Specificity

Detects human NMNAT-1 in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human NMNAT-1 Antibody (675112)

Detection of NMNAT‑1 in Human Kidney.

NMNAT‑1 was detected in immersion fixed paraffin-embedded sections of Human Kidney using Mouse Anti-Human NMNAT‑1 Monoclonal Antibody (Catalog # MAB5865) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei in convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of NMNAT-1 by Immunohistochemistry

Detection of NMNAT-1 by Immunohistochemistry

NAD salvage pathway machinery are reduced in RGC relevant layers of the retina and in the optic nerve head. A NAMPT labelling remains detectable across the whole retina in glaucoma but is significantly reduced in the GCC (n = 11 control eyes, 7 glaucoma eyes). B NMNAT1 is significantly reduced in both the GCC and INL in glaucoma and remains stable in the outer retina (n = 11 control eyes, 7 glaucoma eyes). C NMNAT2 is significantly reduced in the GCC in central retina and is otherwise unchanged in the INL and outer retina (n = 11 control eyes, 7 glaucoma eyes. D–F in the ONH, NAMPT, NMNAT1, and NMNAT2 labelling is significantly reduced in the first 500 µm but remain comparable to controls in the proceeding optic nerve (NAMPT: n = 11 control eyes, 5 glaucoma eyes; NMNAT1: n = 10 control eyes, 5 glaucoma eyes; NMNAT2: n = 8 control eyes, 5 glaucoma eyes). GCC ganglion cell complex, INL inner nuclear layer, OR outer retina, ONH optic nerve head, Ctrl control, Glau glaucoma. Scale bars = 50 µm in A–C, 500 µm in D–E Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36681854), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NMNAT-1 by Immunohistochemistry

Detection of NMNAT-1 by Immunohistochemistry

NAD salvage pathway machinery are reduced in RGC relevant layers of the retina and in the optic nerve head. A NAMPT labelling remains detectable across the whole retina in glaucoma but is significantly reduced in the GCC (n = 11 control eyes, 7 glaucoma eyes). B NMNAT1 is significantly reduced in both the GCC and INL in glaucoma and remains stable in the outer retina (n = 11 control eyes, 7 glaucoma eyes). C NMNAT2 is significantly reduced in the GCC in central retina and is otherwise unchanged in the INL and outer retina (n = 11 control eyes, 7 glaucoma eyes. D–F in the ONH, NAMPT, NMNAT1, and NMNAT2 labelling is significantly reduced in the first 500 µm but remain comparable to controls in the proceeding optic nerve (NAMPT: n = 11 control eyes, 5 glaucoma eyes; NMNAT1: n = 10 control eyes, 5 glaucoma eyes; NMNAT2: n = 8 control eyes, 5 glaucoma eyes). GCC ganglion cell complex, INL inner nuclear layer, OR outer retina, ONH optic nerve head, Ctrl control, Glau glaucoma. Scale bars = 50 µm in A–C, 500 µm in D–E Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36681854), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NMNAT-1 by Immunohistochemistry

Detection of NMNAT-1 by Immunohistochemistry

Labelling of NAD salvage pathway machinery in human retina. A NAMPT labelling is detected across all retinal layers, localizing to both nuclear (haematoxylin + ve) and cytoplasmic (haematoxylin −ve) cellular compartments, with the most intense labeling in the INL (n = 11 eyes). B NMNAT1 labelling is strongest in nuclear layers (GCL, INL, ONL) localizing to nuclei (n = 11 eyes). C NMNAT2 labelling localized to cytoplasmic compartments and was detected across all retinal layers, with the greatest intensity in the GCL, reflecting high expression in RGCs (n = 11 eyes). D In the ONH, NAMPT followed the same trend as in the retina, with labelling localizing to both nuclear and cytoplasmic compartments. Labeling intensity did not vary along the length of the optic nerve (n = 11 eyes). E NMNAT1 again localized only to nuclei in the ONH (glial framework), and its intensity was consistent along the length of the optic nerve (n = 10 eyes). F NMNAT2 conversely, localized only to the axonal compartments of the optic nerve and was significantly greater in the first 500 µm of the ONH (n = 8 eyes), potentially reflecting the density of axons relative to glial framework here, or the need for more NMNAT2 at a critical metabolic portion of the axons. N.B. these data do not allow for direct comparison of abundances of these enzymes given the nature of antibody labelling (efficiency and concentration of the antibodies etc.). RNFL retinal nerve fiber layer, GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer, PS photoreceptor segments. Scale bars = 50 µm in A–C, 500 µm in D–E (overview images) and 50 µm in D–E (inset images) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36681854), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of NMNAT-1 by Immunohistochemistry

Detection of NMNAT-1 by Immunohistochemistry

Labelling of NAD salvage pathway machinery in human retina. A NAMPT labelling is detected across all retinal layers, localizing to both nuclear (haematoxylin + ve) and cytoplasmic (haematoxylin −ve) cellular compartments, with the most intense labeling in the INL (n = 11 eyes). B NMNAT1 labelling is strongest in nuclear layers (GCL, INL, ONL) localizing to nuclei (n = 11 eyes). C NMNAT2 labelling localized to cytoplasmic compartments and was detected across all retinal layers, with the greatest intensity in the GCL, reflecting high expression in RGCs (n = 11 eyes). D In the ONH, NAMPT followed the same trend as in the retina, with labelling localizing to both nuclear and cytoplasmic compartments. Labeling intensity did not vary along the length of the optic nerve (n = 11 eyes). E NMNAT1 again localized only to nuclei in the ONH (glial framework), and its intensity was consistent along the length of the optic nerve (n = 10 eyes). F NMNAT2 conversely, localized only to the axonal compartments of the optic nerve and was significantly greater in the first 500 µm of the ONH (n = 8 eyes), potentially reflecting the density of axons relative to glial framework here, or the need for more NMNAT2 at a critical metabolic portion of the axons. N.B. these data do not allow for direct comparison of abundances of these enzymes given the nature of antibody labelling (efficiency and concentration of the antibodies etc.). RNFL retinal nerve fiber layer, GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer, PS photoreceptor segments. Scale bars = 50 µm in A–C, 500 µm in D–E (overview images) and 50 µm in D–E (inset images) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36681854), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human NMNAT-1 Antibody (675112)

Application
Recommended Usage

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Human Kidney

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: NMNAT-1

NMNAT-1 is expressed in the nuclei of all human tissues, with highest expression in skeletal muscle, heart, kidney, pancreas, and brain (1). The enzyme transfers adenylate from ATP to nicotinamide ribonucleotide or nicotinate ribonucleotide to generate NAD+ or deamido-NAD+, and is an essential enzyme for the production of nuclear NAD+ (2). Nuclear NAD+ is required by poly(ADP-ribose) polymerase 1 (PARP-1), which poly-ADP-ribosylates chromatin in response to DNA strand breaks. NMNAT-1 is known to interact with PARP-1, resulting in its activation, but this interaction with PARP-1 is prevented when NMNAT-1 is phosphorylated at Ser136 (3). Nuclear NAD+ levels are also important for the regulation of SIR2 histone deacetylases (4). A naturally occurring Ube4b/NMNAT-1 chimeric protein is directly involved in slowing the degeneration of injured neurons in mice (5). NMNAT activity is required for the activation of tiazofurin, a drug used to treat leukemia (6). Two other NMNAT enzymes are present in humans. NMNAT-2 is localized in the Golgi complex and cytoplasm, and NMNAT-3 is a mitochondrial enzyme (7).

References

  1. Emanuelli, M. et al. (2001) J. Biol.Chem. 276:406.
  2. Schweiger, M. et al. (2001) FEBS Lett. 492:95.
  3. Berger, F. et al. (2007) Proc. Natl. Acad. Sci. USA 104:3765.
  4. Revollo, J.R. et al. (2004) J. Biol. Chem. 279:50754.
  5. Mack, T.G. et al. (2001) Nature Neurosci. 4:1199.
  6. Boulton, S. et al. (1997) Br. J. Cancer 76:845.
  7. Berger, F. et al. (2005) J. Biol. Chem. 280:36334.

Long Name

Nicotinamide mononucleotide adenylyltransferase-1

Alternate Names

NMN adenylyltransferase 1, NMNAT1, PNAT1

Entrez Gene IDs

64802 (Human)

Gene Symbol

NMNAT1

UniProt

Q9HAN9

Additional NMNAT-1 Products

Product Documents for Human NMNAT-1 Antibody (675112)

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Product Specific Notices for Human NMNAT-1 Antibody (675112)

For research use only

Citations for Human NMNAT-1 Antibody (675112)

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