Recombinant Mouse Anti-fHbp Antibody (JAR41)

CAT#: HPAB-0023-WJ

This product is a mouse monoclonal IgG1 antibody targeting the N-terminal domain of Neisseria meningitidis factor H binding protein (fHbp) variants. This antibody is generated from a human factor H (fH) transgenic BALB/c mouse immunized with a recombinant fHbp ID 1. This antibody is bactericidal when tested in combination with a second non-bactericidal anti-fHbp mAb

Specifications

  • Immunogen
  • Recombinant fHbp ID 1 (variant group 1)
  • Host Species
  • Mouse
  • Derivation
  • Hybridoma
  • Type
  • Mouse IgG1
  • Antibody Isotype
  • IgG1
  • Specificity
  • N. meningitidis fHbp
  • Species Reactivity
  • N. meningitidis
  • Clone
  • JAR41
  • Applications
  • ELISA, FC, DB, Inhib
  • Related Disease
  • Neisseria meningitidis infection

Applications

  • Application Notes
  • The fHbp antibody has been reported in applications of Enzyme-linked Immunosorbent Assay, Western Blot, Flow Cytometry.
    For ELISA: Binding of human FH and anti-FHbp MAbs to purified FHbp was performed as described. Briefly the wells of a microtiter plate were coated with 2 µg/mL of purified recombinant FHbp in PBS. After blocking with PBS containing 0.01% (wt/vol) sodium azide and 1% (wt/vol) BSA, serial dilutions of purified human FH, anti-FHbp MAbs, or a control Penta-His MAb were added and incubated at room temperature for 1 h. Bound FH was detected with sheep anti-FH (1:7,000) and donkey anti-sheep IgG conjugated to alkaline phosphatase (1:5,000). Bound MAbs were detected with goat anti-mouse IgG conjugated to alkaline phosphatase (1:5,000). The ELISAs were developed using paranitrophenyl phosphate substrate (1 mg/mL), and the optical density at 405 nm (OD405 nm) was measured after 30 min at room temperature. Relative binding was calculated as the ratio in the concentration of FH or MAb needed to obtain an OD405 nm of 2.0 for WT versus mutant FHbp. FH and mAb binding was tested in at least three experiments, and the means and 2 SE are shown from two experiments, each performed in triplicate.
    For FC: Binding of the murine mAbs to the surface of live encapsulated bacteria was measured by flow cytometry, which was performed as described previously. The group B test strains were H44/76 (B:15:P1.7,16; ST-32) with fHbp ID 1; a previously described isogenic mutant, which had been engineered to have decreased expression of fHbp ID 1; 8047 (B:2b:P1.5-1, 2-2; ST-8), which expressed fHbp ID 77 in variant group 2; and M1239 (B:14:P1.23,14; ST-41/44), which expressed fHbp ID 28 in variant group 3. The bacterial cells were grown in Mueller Hinton broth culture, harvested by centrifugation, washed, and resuspended in buffer as described elsewhere.

Target

  • Alternative Names
  • FHbp; Fa; Factor H binding protein
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(Creative Biolabs Cat# HPAB-0023-WJ, RRID: AB_3713762)

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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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