NAMPT/PBEF Monoclonal antibody

• Featured Product
• KD/KO Validated

NAMPT/PBEF Monoclonal Antibody for IF, IHC, WB, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat

Applications

WB, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

4

CloneNo.

3D4D8

Cat no : 66385-1-Ig

Synonyms

1110035O14Rik, NAmPRTase, NAMPT, NAMPT/PBEF, PBEF, PBEF1, Pre B cell enhancing factor, VF, VISFATIN

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Validation Data Gallery

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  WB analysis using 66385-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF 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.

• 

  WB analysis using 66385-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF 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.

• 

  WB analysis of human heart using 66385-1-Ig

  human heart tissue were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human skeletal muscle using 66385-1-Ig

  human skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat heart using 66385-1-Ig

  rat heart tissue were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse skeletal muscle using 66385-1-Ig

  mouse skeletal muscle tissue were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Neuro-2a using 66385-1-Ig

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human breast cancer using 66385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:200 (under 10x lens).

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  IHC staining of human breast cancer using 66385-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66385-1-Ig (NAMPT/PBEF antibody at dilution of 1:200 (under 40x lens).

• 

  IF Staining of HeLa using 66385-1-Ig

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using NAMPT/PBEF antibody (66385-1-Ig, Clone: 3D4D8 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HepG2 using 66385-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using NAMPT/PBEF antibody (66385-1-Ig, Clone: 3D4D8 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: LNCaP cells, MCF-7 cells, human heart tissue, human skeletal muscle tissue, rat heart tissue, mouse skeletal muscle tissue, Neuro-2a cells, HeLa cells, HepG2 cells, K-562 cells, HSC-T6 cells, PC-12 cells, NIH/3T3 cells, Jurkat cells, PC-3 cells, HT-29 cells, HCT 116 cells, MOLT-4 cells
• Positive IHC detected in: human breast cancer 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 detected in: HeLa cells, HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:200-1:800

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

• KD/KO: See 1 publications below
• WB: See 4 publications below
• IHC: See 1 publications below

Product Information

66385-1-Ig targets NAMPT/PBEF in WB, IHC, IF, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: NAMPT/PBEF fusion protein Ag2434
• Full Name: nicotinamide phosphoribosyltransferase
• Calculated Molecular Weight: 52 kDa
• Observed Molecular Weight: 52 kDa
• GenBank Accession Number: BC020691
• Gene Symbol: NAMPT
• Gene ID (NCBI): 10135
• RRID: AB_2881761
• 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

Nicotinamide phosphoribosyltransferase(NAMPT) has two usual synonyms termed Visfatin and PBEF. Its primary role is to catalyze the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, an intermediate in the biosynthesis of NAD, which is the rate limiting component in the mammalian NAD biosynthesis pathway. NAMPT is localized in cytoplasm and expressed in large amounts in bone marrow, liver tissue, and muscle tissues. NAMPT inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. NAMPT levels are altered in plasma of patients with type 2 diabetes mellitus (T2DM), and it is now evidenced that NAMPT may plays a role in lipid metabolism.

Publications

Species	Application	Title
human	WB,IHC	J Cell Physiol; Nicotinamide phosphoribosyl transferase regulates cell growth via the Sirt1/P53 signaling pathway and is a prognosis marker in colorectal cancer.; Authors - Jing-Hua Pan ; KD Validated
human	WB	Cell Rep; NAD+ supplement potentiates tumor-killing function by rescuing defective TUB-mediated NAMPT transcription in tumor-infiltrated T cells.; Authors - Yuetong Wang
human	WB	Cell Chem Biol; Addressing the Enzyme-independent tumor-promoting function of NAMPT via PROTAC-mediated degradation; Authors - Xiaotong Zhu
mouse	WB	Int J Mol Sci; Regulation of NAD+/NADH Redox Involves the Protective Effects of Ginsenoside Rb1 against Oxygen-Glucose Deprivation/Reoxygenation-Induced Astrocyte Lesions; Authors - Ying Liu