Human PBEF/Visfatin Antibody

Catalog #: AF4335 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4335
AF4335-SP

Detection of Human PBEF/Visfatin by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human PBEF/Visfatin Antibody Summary

Species Reactivity

Human

Specificity

Detects human PBEF/Visfatin in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross‑reactivity with recombinant rat PBEF is observed, and approximately 25% cross-reactivity with recombinant mouse PBEF is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human PBEF/Visfatin isoform 1
Pro27-His491
Accession # P43490

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.2 µg/mL

See below

Simple Western

2 µg/mL

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human PBEF/Visfatin by Western Blot. Western blot shows lysates of human heart tissue, 293T human embryonic kidney cell line, and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human PBEF/Visfatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4335) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PBEF/Visfatin at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Intracellular Staining by Flow Cytometry

Detection of PBEF/Visfatin antibody in PMA-treated Human PBMCs antibody by Flow Cytometry.

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Detection of PBEF/Visfatin in PMA-treated Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells were treated for 24 hours with 50 ng/mL PMA then stained with Sheep Anti-Human PBEF/Visfatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4335, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Northern-Lights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL011) To facilitate intra-cellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Simple Western

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Detection of Human PBEF/Visfatin by Simple Western^TM. Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line, loaded at 0.2 mg/mL. A specific band was detected for PBEF/Visfatin at approximately 58 kDa (as indicated) using 2 µg/mL of Sheep Anti-Human PBEF/Visfatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4335) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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.

Background: PBEF/Visfatin

PBEF (pre-B cell colony-enhancing factor; also visfatin and nicotinamide phosphoribosyltransferase) is a 52 kDa member of the NAPRTase family of molecules. It functions both intracellularly and extracellularly, where it participates in NAD synthesis and insulin receptor activation, respectively. Human PBEF is 491 amino acids in length and contains no signal sequence. There is at least one alternative splice form that shows a 5 aa substitution for the C-terminal 128 amino acids (aa 364-491). Over aa 27-491, human PBEF shares 96%, 97%, and 96% aa identity with mouse, porcine, and canine PBEF, respectively.

Long Name

Pre-B-Cell Colony-Enhancing Factor 1

Entrez Gene IDs

10135 (Human); 59027 (Mouse); 297508 (Rat)

Alternate Names

EC 2.4.2.12; MGC117256; NAmPRTase; NAMPT; nicotinamide phosphoribosyltransferase; NMPRTase; PBEF; PBEF1; PBEF1110035O14Rik; PBEF1DKFZp666B131; Pre-B cell-enhancing factor; pre-B-cell colony enhancing factor 1; Pre-B-cell colony-enhancing factor 1; VF; Visfatin