Recombinant Anti-Visfatin antibody [EPR21980] (ab236874)

Product name

Anti-Visfatin antibody [EPR21980]
See all Visfatin primary antibodies
Description

Rabbit monoclonal [EPR21980] to Visfatin
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: K562, Raji, HeLa, HT-1080, PC-3, MCF7, HCT 116, C6, PC-12 and Raw 264.7 whole cell lysate; Human fetal kidney lysate. IHC-P: Human bladder cancer and breast cancer tissue; Mouse kidney tissue; Rat liver tissue. ICC/IF: HCT 116 and K562 cells. Flow: K562 and HCT 116 cells. IP: K562 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21980
Isotype

IgG
Research areas

Alternative Versions
- Anti-Visfatin antibody [EPR21980] - BSA and Azide free (ab240589)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Visfatin protein (ab198090)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab236874 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Predicted molecular weight: 55 kDa.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
ICC/IF		1/100.
IP		1/30.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Predicted molecular weight: 55 kDa.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
ICC/IF 1/100.
IP 1/30.
Flow Cyt (Intra) 1/500.

Function

Catalyzes the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, an intermediate in the biosynthesis of NAD. It is the rate limiting component in the mammalian NAD biosynthesis pathway.
Tissue specificity

Expressed in large amounts in bone marrow, liver tissue, and muscle. Also present in heart, placenta, lung, and kidney tissues.
Pathway

Cofactor biosynthesis; NAD(+) biosynthesis; nicotinamide D-ribonucleotide from 5-phospho-alpha-D-ribose 1-diphosphate and nicotinamide: step 1/1.
Sequence similarities

Belongs to the NAPRTase family.
Cellular localization

Cytoplasm.
Target information above from: UniProt accession P43490 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10135 Human
- Entrez Gene: 59027 Mouse
- Entrez Gene: 297508 Rat
- Omim: 608764 Human
- SwissProt: P43490 Human
- SwissProt: Q99KQ4 Mouse
- SwissProt: Q80Z29 Rat
- Unigene: 489615 Human
- Unigene: 202727 Mouse
- Unigene: 203508 Rat
see all
Alternative names
- 1110035O14Rik antibody
- AI314458 antibody
- AI480535 antibody
- DKFZP666B131 antibody
- EC 2.4.2.12 antibody
- MGC117256 antibody
- NAmPRTase antibody
- Nampt antibody
- NAMPT_HUMAN antibody
- Nicotinamide phosphoribosyltransferase antibody
- PBEF 1 antibody
- PBEF antibody
- PBEF1 antibody
- Pre B cell colony enhancing factor 1 antibody
- Pre B cell colony enhancing factor antibody
- Pre B cell enhancing factor antibody
- Pre-B cell-enhancing factor antibody
- Pre-B-cell colony-enhancing factor 1 antibody
- VF antibody
- Visfatin antibody
see all

Western blot - Anti-Visfatin antibody [EPR21980] (ab236874)

All lanes : Anti-Visfatin antibody [EPR21980] (ab236874) at 1/1000 dilution

Lane 1 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg
Lane 2 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 4 : HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate at 20 µg
Lane 5 : PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 6 : Human fetal kidney lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 55 kDa

Blocking/Diluting buffer and concentration: 5% NFDM/TBST

Exposure times:
Lanes 1-2: 26 seconds
Lanes 3-5: 92 seconds
Lane 6: 3 minutes

Lane 6 was developed using a higher sensitivity ECL substrate.

The molecular weight observed is consistent with what has been described in the literature (PMID:25368373, PMID:12782293).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Visfatin using ab236874 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear and cytoplasmic staining in human bladder cancer (PMID:26396920) is observed. Counterstained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling Visfatin (Green) with ab236874 at 1/100 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic and nuclear staining in HCT 116 cell line is observed. The nuclear counterstain is DAPI (Blue). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used as the counterstain (Red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa-Fluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
Immunoprecipitation - Anti-Visfatin antibody [EPR21980] (ab236874)

Visfatin was immunoprecipitated from 0.35 mg of K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab236874 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236874 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1: K562 whole cell lysate 10 μg (input).
Lane 2: ab236874 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236874 in K562 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Flow Cytometry (Intracellular) - Anti-Visfatin antibody [EPR21980] (ab236874)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling Visfatin with ab236874 at 1/500 dilution (Red) compared with the Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-Visfatin antibody [EPR21980] (ab236874)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized K562 (human chronic myelogenous leukemia lymphoblast) cell line labeling Visfatin with ab236874 at 1/500 dilution (Red) compared with the Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Visfatin using ab236874 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear and cytoplasmic staining in rat liver (PMID: 29104634; PMID:24603648) is observed. Counterstained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Visfatin using ab236874 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear and cytoplasmic staining in mouse kidney (PMID: 29104634; PMID:24287278) is observed. Counterstained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Visfatin using ab236874 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear and cytoplasmic staining in human breast cancer (PMID:21784959) is observed. Counterstained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Visfatin antibody [EPR21980] (ab236874)

All lanes : Anti-Visfatin antibody [EPR21980] (ab236874) at 1/1000 dilution

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2 : HCT 116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 3 : HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate at 20 µg
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate at 20 µg
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg
Lane 6 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 55 kDa

Blocking/Diluting buffer and concentration: 5% NFDM/TBST

Exposure time:
Lanes 1-3: 40 seconds
Lanes 4-5: 3 minutes
Lane 6: 15 seconds

The molecular weight observed is consistent with what has been described in the literature (PMID:25368373, PMID:12782293).
Immunocytochemistry/ Immunofluorescence - Anti-Visfatin antibody [EPR21980] (ab236874)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (human chronic myelogenous leukemia lymphoblast) cells labeling Visfatin (Green) with ab236874 at 1/100 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic and nuclear staining in K562 cell line is observed. The nuclear counterstain is DAPI (Blue). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used as the counterstain (Red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa-Fluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
Anti-Visfatin antibody [EPR21980] (ab236874)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab236874? Please let us know so that we can cite the reference in this datasheet.

ab236874 has been referenced in 3 publications.

Kim HN et al. A decrease in NAD+ contributes to the loss of osteoprogenitors and bone mass with aging. NPJ Aging Mech Dis 7:8 (2021). PubMed: 33795658
Ying M et al. Lactate and glutamine support NADPH generation in cancer cells under glucose deprived conditions. Redox Biol 46:102065 (2021). PubMed: 34293554
Zhou L et al. NAMPT/SIRT1 Attenuate Ang II-Induced Vascular Remodeling and Vulnerability to Hypertension by Inhibiting the ROS/MAPK Pathway. Oxid Med Cell Longev 2020:1974265 (2020). PubMed: 33488923

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (3)

Customer reviews and Q&As