Anti-Human RAGE Therapeutic Antibody scFv Fragment (XT-M4) (CAT#: TAB-293CT-S(P))

Recombinant monoclonal antibody specifically binds to RAGE and neutralizes RAGE-associated activities. This antibody can be potentially used for treating RAGE-related disease.

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SPR

Figure 1 3B4 has a high affinity to mouse RAGE while the control M4 does not bind mouse RAGE.

SPR sensorgrams of A. anti-RAGE scFv (3B4) and B. control scFv (M4) using the Langmuir 1:1 binding model.

Kim, H. Y., Wang, X., Kang, R., Tang, D., Boone, B. A., Zeh III, H. J., ... & Edwards, W. B. (2018). RAGE-specific single chain Fv for PET imaging of pancreatic cancer. PloS one, 13(3), e0192821.

IF

Figure 2 Confocal microscopic images of RAGE expressing Panc02 cells.

Live cells were incubated with 3B4-Cy5 and M4-Cy5. After fixation, the nucleus was counterstained with DAPI (Scale bar = 10 μm).

Kim, H. Y., Wang, X., Kang, R., Tang, D., Boone, B. A., Zeh III, H. J., ... & Edwards, W. B. (2018). RAGE-specific single chain Fv for PET imaging of pancreatic cancer. PloS one, 13(3), e0192821.

FC

Figure 3 Flow cytometric analysis with fixed and live Panc02 cells demonstrate antibody-binding.

Cells were stained with 3B4-Cy5 and M4-Cy5 (n ≥ 3, ± SEM, *** p < 0.005 to the untreated cells).

Kim, H. Y., Wang, X., Kang, R., Tang, D., Boone, B. A., Zeh III, H. J., ... & Edwards, W. B. (2018). RAGE-specific single chain Fv for PET imaging of pancreatic cancer. PloS one, 13(3), e0192821.


Specifications

  • Immunogen
  • Human RAGE-Fc
  • Host Species
  • Humanized
  • Type
  • Humanized antibody
  • Specificity
  • Human
  • Species Reactivity
  • Human, mouse
  • Clone
  • XT-M4
  • Applications
  • SPR, IF, FC, WB, ELISA, Neut
  • Related Disease
  • RAGE-related disease

Applications

  • Application Notes
  • The RAGE antibody has been reported in applications of Surface Plasmon Resonance, Immunofluorescence, Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay, Neutralization.
    For Surface Plasmon Resonanc: A Biacore X100 was used for binding kinetic study of purified scFvs. To evaluate the binding affinity of scFvs, recombinant mouse RAGE Fc Chimera was immobilized on a CM5 sensor chip via an amine coupling (~2,000 RU). The carboxyl groups of the reference cell were activated and quenched with aminoethanol. The anti-RAGE-Mab, scFvs and conjugates were diluted in HBS-EP buffer (10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.005% surfactant P20, pH 7.4) and flowed over the surface for 180 s at a rate of 30 μL/min followed by dissociation for 600 seconds. After each sample injection, the surface was regenerated with consecutive injections of 5 μL of 0.3% SDS solution and 3.3 μL 50 mM NaOH. All sensorgrams were double referenced by subtracting the surface effect from the control flow cell and the buffer effect from the blank buffer. The calculated kinetic values ka, kd, and KD were obtained using Biacore X100 Evaluation Software assuming the Langmuir 1:1 binding model. The fitted curves are superimposed on the binding isotherms.
    For Flow Cytometry: Panc02 cells were harvested by trypsinization and fixed with 4% PFA. 5 x 104 cells were used for each labeling condition (n ≥ 3). Cells were incubated with 3B4-Cy5 and M4-Cy5 (2 μg) in the incubation buffer (100 μL) for 30 min on ice. Anti-mouse RAGE Mab (1/100) and Alexa Fluor 488 conjugated anti-mouse antibody (1/200) were sequentially incubated with cells for 30 min on ice. After each incubation, cells were washed with cold PBS three times. The same labeling conditions were used for live cell staining. Flow cytometric analysis was performed on a FACS LSR Fortessa flow cytometer using FACSDiva software provided by the University of Pittsburgh Cancer Institute Flow and Imaging Cytometry core facility, and the data was analyzed using VenturiOne software.

Target

  • Alternative Names
  • RAGE; SCARJ1

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 "AGER"


For research use only. Not intended for any 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

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