Overview

Description:

NAMPT / Visfatin Protein LS-G3729 is a Recombinant Human NAMPT / Visfatin produced in E. coli aa 1-491 with His tag(s). It is low in endotoxin; Less than 1.0 EU/µg protein (determined by LAL method). For Research Use Only

Specifications

Type

Recombinant Protein

Target

NAMPT / Visfatin

Synonyms

Species

Human

Modifications

Unmodified

Conjugations

Unconjugated

Tag

His

Region

aa 1-491

Predicted Molecular Weight

~52kDa (SDS-PAGE)

Expression System

E. coli

Source Species

E. coli

Purification

Greater than 90% by SDS-PAGE

Bio-Activity

Shows adipogenic effects in stimulated differentiating 3T3-L1 cells.

Endotoxin

Less than 1.0 EU/µg protein (determined by LAL method).

Presentation

Lyophilized, Salt free.

Reconstitution

Reconstitute with distilled water.

Storage

Store lyophilized at -20°C for up to 1 year. Once reconstituted store at 4°C for immediate use, or aliquot and store at -20°C for up to 3 months. Avoid freeze-thaw cycles.

Restrictions

For research use only. Intended for use by laboratory professionals.

Guarantee

This protein carries the LSBio 100% Guarantee.

LSBio Guarantee

About NAMPT / Visfatin

Uniprot: P43490 NCBI: NM_005746 NP_005737.1

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Images

Functional Assay

NAMPT / Visfatin Protein - Insulin-mimetic effects on stimulated differentiating 3T3-L1 cells. 10 ug/ml iNampt (human) (rec.) (His) or human insulin was added to differentiating 3T3-L1 cells that had been stimulated with 1 uM dexamethasone and 0.5mM IBMX for 2 days. After 5 days, fat droplets were stained with Oil-Red O.

Insulin-mimetic effects on stimulated differentiating 3T3-L1 cells. 10 ug/ml iNampt (human) (rec.) (His) or human insulin was added to differentiating 3T3-L1 cells that had been stimulated with 1 uM dexamethasone and 0.5mM IBMX for 2 days. After 5 days, fat droplets were stained with Oil-Red O.

Functional Assay

NAMPT / Visfatin Protein - Dimer formation of recombinant Nampt (Visfatin, PBEF) (AG-40A-0018). Purified Nampt (Visfatin, PBEF) was separated by SDS-PAGE and Western blot analysis was performed using rabbit anti-Nampt polyclonal antibody (AG-25A-0025). In the absence of DTT, Nampt (visfatin, PBEF) formed a homodimer, a homomultimer.

Dimer formation of recombinant Nampt (Visfatin, PBEF) (AG-40A-0018). Purified Nampt (Visfatin, PBEF) was separated by SDS-PAGE and Western blot analysis was performed using rabbit anti-Nampt polyclonal antibody (AG-25A-0025). In the absence of DTT, Nampt (visfatin, PBEF) formed a homodimer, a homomultimer.

Functional Assay

NAMPT / Visfatin Protein - Measurement of NAMPT enzymatic activity was performed as described previously [G.C. Elliott, et al.; Anal Biochem 107, 199 (1980)]. The recombinant Nampt was diluted in assay buffer and 10 ul per 50 ul reaction mix were applied in the reaction mix (20 mmol/l Tris-HCl pH 7.4; 2,5 mmol/l ATP; 50 mmol/l NaCl; 12,5 mmol/l MgCl2; 2 mmol/l DTT; 0,5 mmol/l PRPP; 5 uMol/l 14C-nicotinamide) and incubated at 37°C for 1h. The 50 ul reaction mix was transferred into tubes containing 2ml of acetone and afterwards pipetted onto acetone-presoaked glass microfibre filters (GF/A 24 mm). After rinsing with 2 x 1ml acetone, filters were dried, transferred into vials with 6ml scintillation cocktail and radioactivity of 14C-NMN was quantified in a liquid scintillation counter. After subtraction of buffer values as background, cpm were normalized to 10^6 cells and the volume of enzyme preparation (10 ul). Mouse liver lysate at a concentration of 34.5 ug/ml was used as positive control in each assay. The positive control is mouse liver lysate at a concentration of 34,5 ug/ml - normally, which brings the most counts per minute (cpm) (contributed by Antje Garten and Dr. Kiess, University of Leipzig, Germany).

Measurement of NAMPT enzymatic activity was performed as described previously [G.C. Elliott, et al.; Anal Biochem 107, 199 (1980)]. The recombinant Nampt was diluted in assay buffer and 10 ul per 50 ul reaction mix were applied in the reaction mix (20 mmol/l Tris-HCl pH 7.4; 2,5 mmol/l ATP; 50 mmol/l NaCl; 12,5 mmol/l MgCl2; 2 mmol/l DTT; 0,5 mmol/l PRPP; 5 uMol/l 14C-nicotinamide) and incubated at 37°C for 1h. The 50 ul reaction mix was transferred into tubes containing 2ml of acetone and afterwards pipetted onto acetone-presoaked glass microfibre filters (GF/A 24 mm). After rinsing with 2 x 1ml acetone, filters were dried, transferred into vials with 6ml scintillation cocktail and radioactivity of 14C-NMN was quantified in a liquid scintillation counter. After subtraction of buffer values as background, cpm were normalized to 10^6 cells and the volume of enzyme preparation (10 ul). Mouse liver lysate at a concentration of 34.5 ug/ml was used as positive control in each assay. The positive control is mouse liver lysate at a concentration of 34,5 ug/ml - normally, which brings the most counts per minute (cpm) (contributed by Antje Garten and Dr. Kiess, University of Leipzig, Germany).

Functional Assay

Insulin-mimetic effects on stimulated differentiating 3T3-L1 cells. 10 ug/ml iNampt (human) (rec.) (His) or human insulin was added to differentiating 3T3-L1 cells that had been stimulated with 1 uM dexamethasone and 0.5mM IBMX for 2 days. After 5 days, fat droplets were stained with Oil-Red O.

Functional Assay

Dimer formation of recombinant Nampt (Visfatin, PBEF) (AG-40A-0018). Purified Nampt (Visfatin, PBEF) was separated by SDS-PAGE and Western blot analysis was performed using rabbit anti-Nampt polyclonal antibody (AG-25A-0025). In the absence of DTT, Nampt (visfatin, PBEF) formed a homodimer, a homomultimer.

Functional Assay

Measurement of NAMPT enzymatic activity was performed as described previously [G.C. Elliott, et al.; Anal Biochem 107, 199 (1980)]. The recombinant Nampt was diluted in assay buffer and 10 ul per 50 ul reaction mix were applied in the reaction mix (20 mmol/l Tris-HCl pH 7.4; 2,5 mmol/l ATP; 50 mmol/l NaCl; 12,5 mmol/l MgCl2; 2 mmol/l DTT; 0,5 mmol/l PRPP; 5 uMol/l 14C-nicotinamide) and incubated at 37°C for 1h. The 50 ul reaction mix was transferred into tubes containing 2ml of acetone and afterwards pipetted onto acetone-presoaked glass microfibre filters (GF/A 24 mm). After rinsing with 2 x 1ml acetone, filters were dried, transferred into vials with 6ml scintillation cocktail and radioactivity of 14C-NMN was quantified in a liquid scintillation counter. After subtraction of buffer values as background, cpm were normalized to 10^6 cells and the volume of enzyme preparation (10 ul). Mouse liver lysate at a concentration of 34.5 ug/ml was used as positive control in each assay. The positive control is mouse liver lysate at a concentration of 34,5 ug/ml - normally, which brings the most counts per minute (cpm) (contributed by Antje Garten and Dr. Kiess, University of Leipzig, Germany).

Functional Assay

Insulin-mimetic effects on stimulated differentiating 3T3-L1 cells. 10 ug/ml iNampt (human) (rec.) (His) or human insulin was added to differentiating 3T3-L1 cells that had been stimulated with 1 uM dexamethasone and 0.5mM IBMX for 2 days. After 5 days, fat droplets were stained with Oil-Red O.

Functional Assay

Dimer formation of recombinant Nampt (Visfatin, PBEF) (AG-40A-0018). Purified Nampt (Visfatin, PBEF) was separated by SDS-PAGE and Western blot analysis was performed using rabbit anti-Nampt polyclonal antibody (AG-25A-0025). In the absence of DTT, Nampt (visfatin, PBEF) formed a homodimer, a homomultimer.

Functional Assay

Measurement of NAMPT enzymatic activity was performed as described previously [G.C. Elliott, et al.; Anal Biochem 107, 199 (1980)]. The recombinant Nampt was diluted in assay buffer and 10 ul per 50 ul reaction mix were applied in the reaction mix (20 mmol/l Tris-HCl pH 7.4; 2,5 mmol/l ATP; 50 mmol/l NaCl; 12,5 mmol/l MgCl2; 2 mmol/l DTT; 0,5 mmol/l PRPP; 5 uMol/l 14C-nicotinamide) and incubated at 37°C for 1h. The 50 ul reaction mix was transferred into tubes containing 2ml of acetone and afterwards pipetted onto acetone-presoaked glass microfibre filters (GF/A 24 mm). After rinsing with 2 x 1ml acetone, filters were dried, transferred into vials with 6ml scintillation cocktail and radioactivity of 14C-NMN was quantified in a liquid scintillation counter. After subtraction of buffer values as background, cpm were normalized to 10^6 cells and the volume of enzyme preparation (10 ul). Mouse liver lysate at a concentration of 34.5 ug/ml was used as positive control in each assay. The positive control is mouse liver lysate at a concentration of 34,5 ug/ml - normally, which brings the most counts per minute (cpm) (contributed by Antje Garten and Dr. Kiess, University of Leipzig, Germany).

Functional Assay

Insulin-mimetic effects on stimulated differentiating 3T3-L1 cells. 10 ug/ml iNampt (human) (rec.) (His) or human insulin was added to differentiating 3T3-L1 cells that had been stimulated with 1 uM dexamethasone and 0.5mM IBMX for 2 days. After 5 days, fat droplets were stained with Oil-Red O.

Functional Assay

Dimer formation of recombinant Nampt (Visfatin, PBEF) (AG-40A-0018). Purified Nampt (Visfatin, PBEF) was separated by SDS-PAGE and Western blot analysis was performed using rabbit anti-Nampt polyclonal antibody (AG-25A-0025). In the absence of DTT, Nampt (visfatin, PBEF) formed a homodimer, a homomultimer.

Functional Assay

Measurement of NAMPT enzymatic activity was performed as described previously [G.C. Elliott, et al.; Anal Biochem 107, 199 (1980)]. The recombinant Nampt was diluted in assay buffer and 10 ul per 50 ul reaction mix were applied in the reaction mix (20 mmol/l Tris-HCl pH 7.4; 2,5 mmol/l ATP; 50 mmol/l NaCl; 12,5 mmol/l MgCl2; 2 mmol/l DTT; 0,5 mmol/l PRPP; 5 uMol/l 14C-nicotinamide) and incubated at 37°C for 1h. The 50 ul reaction mix was transferred into tubes containing 2ml of acetone and afterwards pipetted onto acetone-presoaked glass microfibre filters (GF/A 24 mm). After rinsing with 2 x 1ml acetone, filters were dried, transferred into vials with 6ml scintillation cocktail and radioactivity of 14C-NMN was quantified in a liquid scintillation counter. After subtraction of buffer values as background, cpm were normalized to 10^6 cells and the volume of enzyme preparation (10 ul). Mouse liver lysate at a concentration of 34.5 ug/ml was used as positive control in each assay. The positive control is mouse liver lysate at a concentration of 34,5 ug/ml - normally, which brings the most counts per minute (cpm) (contributed by Antje Garten and Dr. Kiess, University of Leipzig, Germany).