Anti-Human RAGE Recombinant 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.

Published Data Gene Expression
Figure 1 3B4 has a high affinity to mouse RAGE while the control M4 does not bind mouse RAGE. Figure 2 Confocal microscopic images of RAGE expressing Panc02 cells. Figure 3 Flow cytometric analysis with fixed and live Panc02 cells demonstrate antibody-binding.
Figure 1 IF staining of human cell line MCF7 Figure 2 Lung Figure 3 RNA cell line category: Cell line enhanced (HEK93, JURKAT, RPTEC TERT1, U-266/70, U-266/84)

Specifications

  • Immunogen
  • Human RAGE-Fc
  • Host Species
  • Humanized
  • Type
  • Humanized antibody
  • Specificity
  • Human
  • Species Reactivity
  • Human, mouse
  • Clone
  • XT-M4
  • 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 Resonance: 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 fluorescence 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
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Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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Protocol & Troubleshooting

We have outlined the assay protocols, covering reagents, solutions, procedures, and troubleshooting tips for common issues in order to better assist clients in conducting experiments with our products. View the full list of Protocol & Troubleshooting.

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