Anti-Human CD14 Recombinant Antibody (scFv2F9) (CAT#: TAB-1589CL)
This single chain antibody fragment that specifically binds to CD14 with good positive cells without recognition of other organizations can be used in antiendotoxin therapy.
Specific Inquiry
WB
Figure 1 Expression of scFv2F9 in CHO cells.
(A) SDS-PAGE analysis of the scFv2F9. Lane M: Prestained Protein Molecular Weight Marker, Lane 1: supernatant of the CHO cells transfected with the pSectag2A, Lane 2: Supernatant of the CHO cells transfected with the pSectag2A/ScFv2F9. (B) Western-blot analysis of ScFv2F9. The lane symbols are the same as denoted in (A).
Tang, Y. M., Ning, B. T., Cao, J., Shen, H. Q., & Qian, B. Q. (2007). Construction and expression of single-chain antibody derived from a new clone of monoclonal antibody against human CD14 in CHO cells. Immunopharmacology and immunotoxicology, 29(3-4), 375-386.
FC
Figure 2 The blocking test of ScFv2F9 secreted from CHO cells against 2F9-FITC.
5 groups including background control (using antigen-unrelated murine IgG1 antibody), negative control (using 2F9-FITC), positive control (purified parental 2F9 McAb), culture supernatant from pSectag2A/scFv2F9 and culture supernatant from pSectag2A were designed. 5 ×10⁵ PBMC were added to each tube suspended with 50 μl of PBS. After washing and re-suspending with 500 μl of PBS, fluorescent signals of FITC were detected through FCM. Three parameters as percentage of positive cells (Percentage), mean fluorescence intensity (MFI) and peak channel (PeakCh) were used to evaluate the blocking efficacy.
Tang, Y. M., Ning, B. T., Cao, J., Shen, H. Q., & Qian, B. Q. (2007). Construction and expression of single-chain antibody derived from a new clone of monoclonal antibody against human CD14 in CHO cells. Immunopharmacology and immunotoxicology, 29(3-4), 375-386.
Block
Figure 3 The blocking effect of ScFv2F9 secreted from CHO cells against 2F9-FITC.
Histogram of the blocking effect of ScFv2F9 secreted from CHO cells against 2F9-FITC was shown according to the results of FCM, the ordinate presented blocking percentage, 3 parameters' percentage, peakCh and MFI were used to evaluate the blocking effect of the supernatant from CHO cell transfected with pSectag2A/ScFv2F9, 2F9 McAb and supernatant from CHO cell transfected with pSectag2A.
Tang, Y. M., Ning, B. T., Cao, J., Shen, H. Q., & Qian, B. Q. (2007). Construction and expression of single-chain antibody derived from a new clone of monoclonal antibody against human CD14 in CHO cells. Immunopharmacology and immunotoxicology, 29(3-4), 375-386.
Specifications
- Host Species
- Human
- Type
- Human antibody
- Specificity
- Human
- Clone
- scFv2F9
- Applications
- WB, FC, Block
- Related Disease
- Antiendotoxin therapy
Applications
- Application Notes
- Western blot
SDS-PAGE were performed. with 12% separation polyacrylamide gel and 5% condensed gel to analyze the expressed scFv2F9, then, the gel was stained with Coomassie blue R250. For Western-blot analysis, the scFv2F9 was transferred to PVDF membrane. Blocked with 5% skim milk, the transferred membrane was incubated with 0.5 μg/ml anti-His antibody for 2 h at 37°C The interesting protein band was detected with goat anti-mouse IgG(H+L) conjugated with horseradish peroxidase (HRP).
Flow cytometry
To determine the binding activity, blocking test against 2F9-FITC was carried out using the flow cytometry (FCM). Five groups including background control (using antigen-unrelated murine IgG1 antibody), negative control (using 2F9-FITC), positive control (purified parental 2F9 McAb), culture supernatant from pSectag2A/scFv2F9 and culture supernatant from pSectag2A were designed. Each tube was labeled with 1, 2, 3, 4 and 5. Human peripheral blood mononuclear cells (PBMC) were collected using the lymphocyte separating medium. After wash with phosphate buffer saline (PBS), 5 ×10⁵ PBMC were added to each tube suspended with 50μl of PBS. 2F9 McAb was added to tube 3. Supernatants of CHO cells transfected with either pSectag2A/scFv2F9 or blank vector pSectag2A were added to tube 4 and 5, respectively. After incubation at 4°C protected from light for 30 min, tube 3, 4 and 5 were washed with PBS. Finally, 2.5μl of γ1-FITC was added to tube 1, 2.5μl of 2F9-FITC was added to tube 2, 3, 4 and 5, respectively. Each tube was mixed well and incubated at 4°C protected from light for 30 min. After wash and re-suspended with 500μl of PBS, fluorescent signals of FITC was detected through FCM.
Target
- Alternative Names
- CD14; CD14 molecule; monocyte differentiation antigen CD14; myeloid cell-specific leucine-rich glycoprotein
- Gene ID
- 929
- UniProt ID
- P08571
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 For Clinical Use.
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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