Recombinant Mouse Anti-Pfs25 Antibody (4B7) (CAT#: FAMAB-0226-CN)

4B7 binds P. falciparum surface protein Pfs25, a molecule expressed on the surface of ookinetes, and inhibits parasite development completely when fed to Anopheles mosquitoes in a gametocytemic bloodmeal.


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ELISA

Figure 1 Competition ELISA with mouse monoclonal antibodies against rabbit, monkey, or human anti-Pfs25 serum.

Figure 1 Competition ELISA with mouse monoclonal antibodies against rabbit, monkey, or human anti-Pfs25 serum.

A fixed amount of mouse monoclonal anti-Pfs25 antibody 4B7 was tested with 2-fold dilutions of anti-Pfs25 serum from rabbit, monkey and humans. The amount of Pfs25-specific antibodies in the diluted serum (competitor) is shown on the x-axis and the O.D. value from the individual mouse monoclonal antibody is shown on the y-axis. The mean and standard deviation were calculated from two independent experiments. For some data points, the standard deviations are too small and are not visible.

Cheru, L., Wu, Y., Diouf, A., Moretz, S. E., Muratova, O. V., Song, G., ... & Miura, K. (2010). The IC50 of anti-Pfs25 antibody in membrane-feeding assay varies among species. Vaccine, 28(27), 4423-4429.

IB

Figure 2 Immunoblot of reduced and non-reduced algae-produced Pfs25 with monoclonal transmission blocking antibodies.

Figure 2 Immunoblot of reduced and non-reduced algae-produced Pfs25 with monoclonal transmission blocking antibodies.

Reduced and non-reduced a-Pfs25 was resolved by SDS-PAGE, transferred to nitrocellulose, anddetected with anti-Pfs25 4B7 mAb.

Gregory, J. A., Li, F., Tomosada, L. M., Cox, C. J., Topol, A. B., Vinetz, J. M., & Mayfield, S. (2012). Algae-produced Pfs25 elicits antibodies that inhibit malaria transmission. PloS one, 7(5), e37179.

WB

Figure 3 Western blot analysis of P.falciparum mixed sexual stage lysates with anti-Pfs25-4B7 mAbs.

Figure 3 Western blot analysis of P.falciparum mixed sexual stage lysates with anti-Pfs25-4B7 mAbs.

Reduced and non-reduced sexual stagelysates were resolved by SDS-PAGE and transferred to nitrocellulose. Blots were probed with anti-Pfs25-4B7 mAbs. (R-reduced, N-non-reduced)

Gregory, J. A., Li, F., Tomosada, L. M., Cox, C. J., Topol, A. B., Vinetz, J. M., & Mayfield, S. (2012). Algae-produced Pfs25 elicits antibodies that inhibit malaria transmission. PloS one, 7(5), e37179.

IF

Figure 4 Indirect immunofluorescence using immune sera from mice immunized with algae-produced Pfs25 on in-vitro cultured P. falciparum gametocytes, gametes, and zygotes.

Figure 4 Indirect immunofluorescence using immune sera from mice immunized with algae-produced Pfs25 on in-vitro cultured P. falciparum gametocytes, gametes, and zygotes.

DNA was stained using DAPI (blue) and antibody binding was visualized using Alexa Fluor 488-conjugated rabbit anti-mouse IgG (green) for anti-Pfs25 4B7 mAbs. Scale bars, 1 μm.

Gregory, J. A., Li, F., Tomosada, L. M., Cox, C. J., Topol, A. B., Vinetz, J. M., & Mayfield, S. (2012). Algae-produced Pfs25 elicits antibodies that inhibit malaria transmission. PloS one, 7(5), e37179.


Specifications

  • Host Species
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • P. falciparum Pfs25
  • Species Reactivity
  • P. falciparum
  • Clone
  • 4B7
  • Applications
  • ELISA, IB, WB, IF

Product Property

  • Storage
  • Store at -20°C for long-term storage. Avoid freeze/thaw cycles.

Applications

  • Application Notes
  • This antibody has been reported for use in Enzyme-Linked Immunosorbent Assay, Immunoblot, Western Blot and Immunofluorescence.
    Western Blot: Plasmodium falciparum strain NF54 was maintained in vitro incontinuous cultivation. Gametocytes, macrogametes, zygotes, andookinetes were cultured and purified as previously described. For antibodies raised to a-Pfs25, 2 mg of reduced and nonreducedparasite lysate was prepared as described above and resolved on RUNBLUE 12% SDS-PAGE precast gels. Resolved proteins were transferred to nitrocellulose and probed withantisera raised to a-Pfs25 at a 1:100 dilution. Identical blots were prepared and probed with anti-Pfs25 4B7 mAbs or preimmunesera. All membranes were visualized using AP-conjugated goat anti-mouse IgG antibodies.
    Immunofluorescence: Fixed, permeabilized gametocytes, macrogametes, and zygoteswere subjected to IFA. Parasites were heat-fixed onto 10-well glassslides. Fixed cells were permeabilized byincubation in PBS containing 3% bovine serum albumin and 0.1% Triton X-100 at room temperature for 15 minutes followed by blocking for 2 hrs at room temperature in 3% BSA in PBS-T. The preparations were incubated with a-Pfs25 antisera (1:100 dilution) or 4b7 mAbs overnight at 4uC followed by AlexaFluor 488 goat anti-mouse IgG (1:500 dilution)at room temperature for 1 hr. Parasite nuclei were stained with 2mg/mL 49-6-Diamidino-2-phenylindole (DAPI) at room temperature for 5 min. Slides were mountedwith Slow FadeHanti-fade kit and images were acquired using an Applied Precision Spectris microscope and deconvolved using Softworx software. Images were adjusted for contrast in Softworx and exported as tiffs and assembled with Adobe Photoshop.

Target

  • Alternative Names
  • Pfs25

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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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.

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