100 µG

PRODUCT NO. HYB 211-01

Anti-NGAL (human, neutrophil gelatinase-associated lipocalin)

PRESENTATION

Preparation: Protein-A purified

Content: Available in 200 µL and 1 mL volumes, 1 mg/mL
Solvent: 0.01 M phosphate buffer, pH 7.4, containing 0.5 M NaCl and 15 mM sodium azide
Storage: In the dark at 4-8ºC

ANTIGEN
Neutrophil gelatinase-associated lipocalin (NGAL; also called lipocalin 2, siderocalin and neutrophil lipocalin) is a member of the lipocalin family of proteins which bind and transport small lipophilic molecules. NGAL is released by activated neutrophils and occurs as 25-kDa glycosylated single protein chain monomers, which form dimers and small amounts of higher oligomers, as well as complexes with matrix metalloproteinase 9 (MMP-9; gelatinase B) (1). Low level expression of NGAL in a variety of epithelia may be increased in inflammation or cancers (2). The
expression and release of NGAL from renal tubules are dramatically increased by ischemic or nephrotoxic injury (3).

IMMUNOGEN

 Human recombinant NGAL in monomer and dimer forms adsorbed to aluminium hydroxide gel (4).

SPECIFICITY HYB

 211-01 is specific for human NGAL. No cross-reaction with NGAL from other species has been detected.

EPITOPE SPECIFICITY
Mouse monoclonal antibody IgG1/k

REACTIVITY
HYB 211-01 reacts strongly with human NGAL. Strong reaction is seen when used as detection antibody insandwich ELISA in combination with a polyclonal NGAL capture antibody and when tested on recombinant NGAL coated directly onto the well. HYB 211-01 reacts with NGAL in immunochemical staining of paraformaldehyde-fixed sections containing neutrophils. In Western blotting of SDS-PAGE gels, HYB 211-01 reacts with NGAL in both reduced and unreduced forms (4).

CULTURE MEDIUM

 RPMI 1640 with 10% fetal calf serum

FUSION PARTNER X63-Ag8.653

IMMUNIZATION

 Female CF1 x BALB/c mice immunized by intraperitoneal injection




APPLICATION

Method                                 Usability         Dilution guideline           References

ELISA                                   Yes                      1/16,000                     4,3
Immunoblotting                       Yes                      1/500                          4
Immunohistochemistry             Yes                                                         4

1. Kjeldsen L, Johnsen AH, Sengeløv H, Borregaard N (1993) Isolation and primary structure of NGAL, a novel protein
associated with human neutrophil gelatinase. J Biol Chem 268:10425-10432.
2. Nielsen BS, Borregaard N, Bundgaard JR, Timshel S, Sehested M, Kjeldsen L (1996) Induction of NGAL synthesis in
epithelial cells of human colorectal neoplasia and inflammatory bowel diseases. Gut 38:414-420.
3. Mishra J, Ma Q, Prada A, Mitsnefes M, Zahedi K, Yang J, Barasch J, Devarajan P (2003) Identification of neutrophil
gelatinase-associated lipocalin as a novel early urinary biomarker for ischemic renal injury. J Am Soc Nephrol 14:2534-2543.
4. Kjeldsen L, Koch C, Arnljots K, Borregaard N (1996) Characterization of two ELISAs for NGAL, a newly described
lipocalin in human neutrophils. J Immunol Methods 198:155-164.
Further examples of use, please consult NGAL reference booklet at www.bioporto.com
REFERENCES
Usability Dilution guideline References
The dilution guideline for ELISA is based on use as detection antibody for antigen coated at 0.5 µg/ml. Users should determine the optimal dilutions for their own