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Human PBEF/Visfatin Antibody

R&D Systems, part of Bio-Techne | Catalog # AF4335

R&D Systems, part of Bio-Techne
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AF4335
AF4335-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

CyTOF-ready, Intracellular Staining by Flow Cytometry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

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

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human PBEF/Visfatin Antibody

Detection of Human PBEF/Visfatin antibody by Western Blot.

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

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.
Detection of Human PBEF/Visfatin antibody by Simple WesternTM.

Detection of Human PBEF/Visfatin by Simple WesternTM.

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.

Applications for Human PBEF/Visfatin Antibody

Application
Recommended Usage

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
Sample: Human peripheral blood mononuclear cells treated with PMA, fixed with paraformaldehyde, and permeabilized with saponin

Simple Western

2 µg/mL
Sample: HEK293T human embryonic kidney cell line

Western Blot

0.2 µg/mL
Sample:   Human heart tissue, 293T human embryonic kidney cell line, and A431 human epithelial carcinoma cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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.

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

Alternate Names

NAmPRTase, NAMPT, NMPRTase, PBEF1, Visfatin

Entrez Gene IDs

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

Gene Symbol

NAMPT

UniProt

Additional PBEF/Visfatin Products

Product Documents for Human PBEF/Visfatin Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human PBEF/Visfatin Antibody

For research use only

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