Mouse Anti-LEP Recombinant Antibody (clone 9F8) (CAT#: PABL-259)

Recombinant Mouse Antibody (9F8) is capable of binding to Leptin. This antibody blocks leptin binding through only a small (10%) overlap in their binding sites, and that leptin binding is likely to involve an induced fit mechanism.

Specific Inquiry
  • Size:
  • Conjugation:
  • Endotoxin:
  • Purity:
  • Fc Engineering:
  • Published Data
  • Datasheet
  • MSDS
  • COA

Figure 1 Competitive binding of leptin and 9F8 to ObR. The binding of leptin-biotin to the full-length extracellular domain of ObR was measured in the presence of either 9F8 mAb or unlabeled leptin. The IC50 for leptin and 9F8 mAb binding to ObR are approximately 0.76 nM and 1.0 nM, respectively.

Carpenter, B., Hemsworth, G. R., Wu, Z., Maamra, M., Strasburger, C. J., Ross, R. J., & Artymiuk, P. J. (2012). Structure of the human obesity receptor leptin-binding domain reveals the mechanism of leptin antagonism by a monoclonal antibody. Structure, 20(3), 487-497.

Figure 2 The affinity of leptin binding to recombinant LBD was calculated by competitive binding assay (data shown represent the mean of three independent experiments, and error bars indicate the standard deviation between data sets). The IC50 of leptin binding to LBD is 0.78 ± 0.05 nM (SD).

Carpenter, B., Hemsworth, G. R., Wu, Z., Maamra, M., Strasburger, C. J., Ross, R. J., & Artymiuk, P. J. (2012). Structure of the human obesity receptor leptin-binding domain reveals the mechanism of leptin antagonism by a monoclonal antibody. Structure, 20(3), 487-497.

Figure 3 Gel filtration analysis of leptin and 9F8 Fab binding to LBD.

Leptin, LBD, and 9F8 Fab each resolve as a single predominant peak with retention volumes of 16.72, 15.94, and 14.95 ml, respectively (peaks shown in parentheses represent contaminants in the protein preparations). 1:1 molar ratio mixtures of LBD/leptin and LBD/9F8 Fab resolve with retention volumes of 15.32 and 13.99 ml, respectively. A 1:1:1 molar ratio mixture of all three proteins resolves as a predominant peak with a retention volume of 14.04 ml, which relates well to the LBD-9F8 Fab complex (13.99 ml). The shoulder peak (approximately 15.35 ml) is likely to contain both free 9F8 Fab (14.95 ml) and the LBD-leptin complex (15.32 ml). Significantly, no peak with a retention volume lower than 14.04 ml was present, demonstrating that leptin and 9F8 Fab cannot simultaneously bind to LBD.

Carpenter, B., Hemsworth, G. R., Wu, Z., Maamra, M., Strasburger, C. J., Ross, R. J., & Artymiuk, P. J. (2012). Structure of the human obesity receptor leptin-binding domain reveals the mechanism of leptin antagonism by a monoclonal antibody. Structure, 20(3), 487-497.

Figure 4 The mAb 2H6 or mAb 9F8, respectively, were immobilized on microtiter plates.

Binding of the biotinylated soluble leptin receptor to mAb 9F can be displaced by leptin, whereas binding to mAb 2H6 is independent from leptin. Antisoluble leptin receptor mAb 2H6 binds to a protein in human serum, which is able to bind biotinylated leptin. The binding of this soluble receptor to biotinylated human leptin is displaceable by nonlabeled human, mouse, and rat leptin but not by nonlabeled hGH or IL-6.

Wu, Z., Bidlingmaier, M., Liu, C., De Souza, E. B., Tschöp, M., Morrison, K. M., & Strasburger, C. J. (2002). Quantification of the soluble leptin receptor in human blood by ligand-mediated immunofunctional assay. The Journal of Clinical Endocrinology & Metabolism, 87(6), 2931-2939.


Specifications

  • Immunogen
  • Human leptin
  • Host Species
  • Mouse
  • Derivation
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • Tested positive against Human LEP
  • Species Reactivity
  • Human
  • Clone
  • 9F8
  • Applications
  • WB, ELISA, FuncS

Target

  • Alternative Names
  • LEP; leptin; OB; OBS; LEPD; obese protein; obesity factor; obese; mouse; homolog of; leptin (murine obesity homolog); leptin (obesity homolog; mouse)

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

See other products for "LEP"

See other products for "Clone 9F8"


For Research Use Only. Not For Clinical Use.

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

Send Inquiry

© 2021 Creative BioLabs. All Rights Reserved
  • 0
  • 0

Cart