NME2 Antikörper (C-Term)

Produktnummer ABIN3044198

Produktdetails anti-NME2 Antikörper

Target: NME2 (Nucleoside diphosphate kinase B (NME2))

Bindungsspezifität: AA 137-152, C-Term

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser NME2 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC)

Verwendungszweck: Rabbit IgG polyclonal antibody for Nucleoside diphosphate kinase B(NME2) detection. Tested with WB, IHC-P, ICC in Human,Mouse,Rat.

Sequenz: EELVDYKSCA HDWVYE

Kreuzreaktivität (Details): No cross reactivity with other proteins.

Produktmerkmale: Rabbit IgG polyclonal antibody for Nucleoside diphosphate kinase B(NME2) detection. Tested with WB, IHC-P, ICC in Human,Mouse,Rat.
Gene Name: NME/NM23 nucleoside diphosphate kinase 2
Protein Name: Nucleoside diphosphate kinase B(NDK B)

Aufreinigung: Immunogen affinity purified.

Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human NME2(137-152aa EELVDYKSCAHDWVYE), different from the related mouse and rat sequences by one amino acid.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB: Concentration: 0.1-0.5 μg/mL, Tested Species: Human, Rat, Predicted Species: Mouse
IHC-P: Concentration: 0.5-1 μg/mL, Tested Species: Human, Mouse, Rat, Epitope Retrieval by Heat: Boiling the paraffin sections in 10 mM citrate buffer, pH 6.0, for 20 mins is required for the staining of formalin/paraffin sections.
ICC: Concentration: 0.5-1 μg/mL, Tested Species: Human
Notes: Tested Species: Species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities. Other applications have not been tested. Optimal dilutions should be determined by end users.

Kommentare:

Antibody can be supported by chemiluminescence kit ABIN921124 in WB, supported by ABIN921231 in IHC(P) and ICC.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #300030 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300030

Datum: 13.12.2016

Antigen: P18

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: tonsil, urinary bladder

Bewertung: Passed. ABIN3044198 staining is consistent with staining pattern of other commercial P18/NME2 antibodies.

Validierungsbilder

Staining of human tonsil (A.) and urinary bladder (B.) sections in a tissue microarray with ABIN3044198. See the protocol for detailed information.

Protokoll

Primärantikörper: ABIN3044198

Sekundärantikörper: HRP polymer (ThermoFisher Scientific, TL-125-PH)

Full Protocol:

• Sample preparation:
• • Fixation and TMA preparation according to PMID: 22688270.
  • Cut 4µm TMA sections using a waterfall microtome (ThermoFisher Scientific, HM355S).
  • Dry sections ON at RT and then back them for 12-24h at 50°C.
• Deparaffinization and rehydration in xylene and graded alcohol series:
• • xylene twice for 3min.
  • 100% ethanol for 1min.
  • 95% ethanol for 1min.
  • 0.3% H₂O₂ in 95% ethanol for 5min to block endogenous peroxidase.
  • 70% ethanol for 1min.
  • 50% ethanol for 1min.
  • distilled H₂O for 2min.
• Antigen Retrieval by Heat Induced Epitope Retrieval (HIER):
• • Boil the sections in Lab Vision Citrate buffer pH6.0 (ThermoFisher Scientific, TA-250-PM1X) for 4min at 125°C in a decloaking chamber (Biocare Medical).
  • Allow slides to cool down to 90°C in the decloaking chamber.
• Immunostaining in a Lab Vision Autostainer 480 (ThermorFisher Scientific) at RT with 300µl of reagents for each step:
• • Rinse sections in Lab Vision wash buffer with extra tween added to a final concentration of 0.2% (ThermoFisher Scientific, TA-999-TT and TA-125-TW) wash buffer.
  • Block sections with Lab Vision Ultra V-Block (ThermoFisher Scientific, TA-125-UB) for 5min.
  • Rinse sections 2x in wash buffer.
  • Incubate sections with primary rabbit anti-P18/NME2 antibody (antibodies-online, ABIN3044198) diluted 1:25000 for 30min.
  • Rinse sections 3x in wash buffer.
  • Incubate sections with labeled HRP polymer (ThermoFisher Scientific, TL-125-PH) for 30min.
  • Rinse sections 2x in wash buffer.
  • Develop in DAB Quanto solution (ThermoFisher Scientific, TL-125-QHDX) for 5min.
  • Rinse sections in distilled H₂O.
• Washing, counterstaining, and coverslipping in Autostainer XL (Leica Biosystems, ST5010) at RT:
• • Counterstaining in Mayer’s hematoxylin plus (HistoLab, 01820) for 7.5min.
  • Rinse sections in tap water for 5min.
  • Rinse sections in lithium carbonate water diluted 1:5 for 5min.
  • Dehydration in graded ethanol series and Neo-Clear (Merck Millipore, 109843) according to the manufacturer recommendations.
  • Automated mounting (Leica Biosystems, CV5030) of coverslip with Pertex (Histolab, 00871).
• Microscopy:
• • Image acquisition on a Leica Aperio AT2 and Aperio Scanscope AT at 20x magnification.

Anmerkungen: ABIN3044198 shows staining in most tissues similar to alternative P18/NME2 antibodies. Determination of the appropriate dilution factor proved difficult.

Handhabung

Format: Lyophilized

Rekonstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Konzentration: 500 μg/mL

Buffer: Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Thimerosal, 0.05 mg Sodium azide.

Konservierungsmittel: Thimerosal (Merthiolate), Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide and Thimerosal (Merthiolate): POISONOUS AND HAZARDOUS SUBSTANCES which should be handled by trained staff only.

Handhabung: Avoid repeated freezing and thawing.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20 °C for a longer time. Avoid repeated freezing and thawing.

Haltbarkeit: 12 months

Details zu NME2

Target: NME2 (Nucleoside diphosphate kinase B (NME2))

Andere Bezeichnung: NME2 (NME2 Produkte)

Synonyme: NDKB antikoerper, NDPK-B antikoerper, NDPKB antikoerper, NM23-H2 antikoerper, NM23B antikoerper, PUF antikoerper, nm23-2 antikoerper, p18-12d antikoerper, nm23-M2 antikoerper, NM23-C2 antikoerper, NME/NM23 nucleoside diphosphate kinase 2 antikoerper, non-metastatic cells 2, protein (NM23B) expressed in antikoerper, NME2 antikoerper, Nme2 antikoerper

Hintergrund: NME2(NME/NM23 nucleoside diphosphate kinase 2) also called non-metastatic cells 2, protein(NM23B) expressed in, NM23-H2, NM23B or NDPKB, is identical to the beta subunit of human erythrocyte NDP kinase. Little to no expression was detected in other mouse tissues examined. Using Northern blot analysis, Masse et al.(2002) detected high expression of mouse Nme2, which they called nm23-M2, in heart, liver, and kidney, with intermediate expression in skeletal muscle. In situ hybridization of 15-day postcoitum mouse embryos showed ubiquitous Nme2 expression. The mouse and human NME2 genes contain 5 exons and span about 6.0 kb. The NME2 gene is mapped on 17q21.33. Srivastava et al.(2006) concluded that histidine phosphorylation regulates KCa3.1 channel activity and that NDPBK is critical to the channel activity and the activation of CD4 T cells.

Synonyms: C myc purine binding transcription factor PUF antibody|C myc transcription factor antibody|C-myc purine-binding transcription factor PUF antibody|Histidine protein kinase NDKB antibody|MGC111212 antibody|MGC2212 antibody|NDK B antibody|NDKB antibody|NDKB_HUMAN antibody|NDP kinase B antibody|NDPK B antibody|NDPKB antibody|NM23 H2 antibody|nm23-H2 antibody|NM23B antibody|nme2 antibody|Non metastatic cells 2, protein(NM23B) expressed in antibody|Nucleoside diphosphate kinase B antibody|Nucleotide diphosphate kinase B antibody|PUF antibody

UniProt: P22392