NAPRT1 Monoclonal antibody

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• KD/KO Validated

NAPRT1 Monoclonal Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

Publications

6

CloneNo.

5D8H10

Cat no : 66159-1-Ig

Synonyms

FHA HIT interacting protein, FHIP, NAPRT1, NAPRTase, PP3856

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Validation Data Gallery

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  WB analysis using 66159-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66159-1-Ig (NAPRT1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

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  WB analysis using 66159-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66159-1-Ig (NAPRT1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 cells using 66159-1-Ig

  WB result of NAPRT1 antibody (66159-1-Ig, 1:6000) with si-Control and si-NAPRT1 transfected HepG2 cells.

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  WB analysis of HEK-293 using 66159-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66159-1-Ig (NAPRT1 Antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 66159-1-Ig

  HepG2 cells were subjected to SDS PAGE followed by western blot with 66159-1-Ig (NAPRT1 Antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  IHC staining of human small intestine using 66159-1-Ig

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 66159-1-Ig (NAPRT1 Antibody) at dilution of 1:50 (under 10x lens).

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  IHC staining of human small intestine using 66159-1-Ig

  Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 66159-1-Ig (NAPRT1 Antibody) at dilution of 1:50 (under 40x lens).

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  IF Staining of HeLa using 66159-1-Ig

  Immunofluorescent analysis of (-20℃ Ethanol ) fixed HeLa cells using 66159-1-Ig(NAPRT1 antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of HeLa using 66159-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using NAPRT1 antibody (66159-1-Ig, Clone: 5D8H10 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: THP-1 cells, HEK-293 cells, Jurkat cells, HepG2 cells, LNCaP cells, HeLa cells, K-562 cells
• Positive IHC detected in: human small intestine tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:20-1:200
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 6 publications below

Product Information

66159-1-Ig targets NAPRT1 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: NAPRT1 fusion protein Ag4265
• Full Name: nicotinate phosphoribosyltransferase domain containing 1
• Calculated Molecular Weight: 514 aa, 55 kDa
• Observed Molecular Weight: 51 kDa
• GenBank Accession Number: BC032466
• Gene Symbol: NAPRT1
• Gene ID (NCBI): 93100
• RRID: AB_2881555
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Nicotinic acid (NA) is a coenzyme in cellular redox reactions, and is an essential component of metabolic pathways in all living cells. NAPRT1 (Nicotinate phosphoribosyltransferase) is essential for increasing cellular NAD levels and, thus, to prevent oxidative stress of cells. NAPRT1 converts Nicotinic acid (NA; niacin) to NA mononucleotide (NaMN), which is then converted to NA adenine dinucleotide (NaAD), and finally to nicotinamide adenine dinucleotide (NAD).

Publications

Species	Application	Title
human	WB	Nat Commun; Extracellular nicotinate phosphoribosyltransferase binds Toll like receptor 4 and mediates inflammation.; Authors - Antonella Managò
human	WB	Nat Commun; PPM1D mutations silence NAPRT gene expression and confer NAMPT inhibitor sensitivity in glioma.; Authors - Nathan R Fons
human	WB	Cell Chem Biol; NAMPT-targeting PROTAC and nicotinic acid co-administration elicit safe and robust anti-tumor efficacy in NAPRT-deficient pan-cancers; Authors - Xiaotong Zhu
human	WB	Mol Cancer Ther; Exploiting Metabolic Defects in Glioma with Nanoparticle Encapsulated NAMPT Inhibitors; Authors - Matthew A Murray
human	WB	Mol Cancer Res; NAPRT silencing in FH-deficient renal cell carcinoma confers therapeutic vulnerabilities via NAD+ depletion; Authors - Katelyn J Noronha
human	WB	Eur J Med Chem; Discovery of potent and novel dual NAMPT/BRD4 inhibitors for efficient treatment of hepatocellular carcinoma; Authors - Chunjia Yin