Recombinant Human Anti-IAV HA Antibody (FI6V3)
CAT#: PABL-214
Recombinant Human Antibody (FI6V3) is capable of binding to IAV HA, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-IAV HA mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-IAV HA mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.
Published Data
Tested Data
Figure 1 IC50 measurements with pseudo virus-based microneutralization assay for the IgGs reformatted from the selected anti-HA trimer scFvs.
Chen, C., Chiu, Y. K., Yu, C. M., Lee, C. C., Tung, C. P., Tsou, Y. L.,... & Yang, A. S. (2017). High throughput discovery of influenza virus neutralizing antibodies from phage-displayed synthetic antibody libraries. Scientific reports, 7(1), 14455.
Figure 2 EC50 measurements with ELISA for the IgGs reformatted from the selected anti-HA trimer scFvs.
Chen, C., Chiu, Y. K., Yu, C. M., Lee, C. C., Tung, C. P., Tsou, Y. L.,... & Yang, A. S. (2017). High throughput discovery of influenza virus neutralizing antibodies from phage-displayed synthetic antibody libraries. Scientific reports, 7(1), 14455.
Figure 3 IC50 measurements with pseudo virus-based microneutralization assay for the IgGs reformatted from the selected anti-HA trimer scFvs.
Chen, C., Chiu, Y. K., Yu, C. M., Lee, C. C., Tung, C. P., Tsou, Y. L.,... & Yang, A. S. (2017). High throughput discovery of influenza virus neutralizing antibodies from phage-displayed synthetic antibody libraries. Scientific reports, 7(1), 14455.
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SDS-PAGE
Figure 1 Recombinant Human Anti-IAV HA Antibody (PABL-214) in SDS-PAGE
SDS-PAGE analysis of PABL-214 in β-mercaptoethanol-reduced (lane 1) and non-reduced (lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 180 kDa.
SEC-HPLC
Figure 2 Recombinant Human Anti-IAV HA Antibody (PABL-214) in SEC-HPLC
The purity of PABL-214 was greater than 93% as determined by SEC-HPLC.
Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl
ELISA
Figure 3 Anti-IAV HA Recombinant Antibody (PABL-214) in ELISA
ELISA analysis of PABL-214 was performed by coating with Influenza A H3N2 (A/Perth/16/2009) Hemagglutinin / HA Protein (His Tag). Then blocked with BSA and incubated with PABL-214. The HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
ELISA
Figure 4 Anti-IAV HA Recombinant Antibody (PABL-214) in ELISA
ELISA analysis of PABL-214 was performed by coating with Influenza A H2N2 (A/Japan/305/1957) Hemagglutinin / HA Protein (His Tag). Then blocked with BSA and incubated with PABL-214. The HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
ELISA
Figure 5 Anti-IAV HA Recombinant Antibody (PABL-214) in ELISA
ELISA analysis of PABL-214 was performed by coating with Influenza A [A/Hong Kong/483/97 (H5N1)] Hemagglutinin (HA) Protein (His Tag). Then blocked with BSA and incubated with PABL-214. The HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
WB
Figure 6 Anti-IAV HA Recombinant Antibody (PABL-214) in WB
Western blot analysis of PABL-214 was performed with Influenza A H3N2 (A/Perth/16/2009) Hemagglutinin / HA Protein (His Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-214 and HRP goat anti-human IgG as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.8 μg)
WB
Figure 7 Anti-IAV HA Recombinant Antibody (PABL-214) in WB
Western blot analysis of PABL-214 was performed with Influenza A H2N2 (A/Japan/305/1957) Hemagglutinin / HA Protein (His Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-214 and HRP goat anti-human IgG as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.8 μg)
WB
Figure 8 Anti-IAV HA Recombinant Antibody (PABL-214) in WB
Western blot analysis of PABL-214 was performed with Influenza A [A/Hong Kong/483/97 (H5N1)] Hemagglutinin (HA) Protein (His Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-214 and HRP goat anti-human IgG as a secondary antibody (1: 2000). Chemiluminescent detection was performed.
Lane 1: Reduced antigen (0.8 μg)
DB
Figure 9 Anti-IAV HA Recombinant Antibody (PABL-214) in DB
Dot Blot analysis of PABL-214 was performed by coating with Influenza A H3N2 (A/Perth/16/2009) Hemagglutinin / HA Protein (His Tag).
PABL-214 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000)
DB
Figure 10 Anti-IAV HA Recombinant Antibody (PABL-214) in DB
Dot Blot analysis of PABL-214 was performed by coating with Influenza A H2N2 (A/Japan/305/1957) Hemagglutinin / HA Protein (His Tag).
PABL-214 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000)
DB
Figure 11 Anti-IAV HA Recombinant Antibody (PABL-214) in DB
Dot Blot analysis of PABL-214 was performed by coating with Influenza A [A/Hong Kong/483/97 (H5N1)] Hemagglutinin (HA) Protein (His tag).
PABL-214 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-human IgG as a secondary antibody (1: 2000)
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Specifications
- Immunogen
- Influenza a virus Hemagglutinin
- Host Species
- Human
- Derivation
- Human
- Type
- IgG
- Specificity
- Tested positive against native IAV HA
- Species Reactivity
- IAV
- Clone
- FI6V3
- Applications
- ELISA, Neut, WB, IHC, FuncS
Product Property
- Purity
- >95% by SDS-PAGE and HPLC analysis
- Storage
- Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing of samples.
Applications
- Application Notes
- The antibody was validated for ELISA, Neutralization. For details, refer to Published Data.
Target
- Alternative Names
- HA; hemagglutinin; haemagglutinin
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Customer Review
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Anna Thompson14.Sep,23
Excellent for Virus Detection
I've been using the Anti-IAV-HA Antibody for detecting influenza A virus in our lab, and it has delivered excellent results. The specificity and sensitivity are top-notch, providing clear and reliable detection. This antibody has significantly improved our virus detection assays and saved us a lot of troubleshooting time.
David Brown05.Dec,22
Reliable for Immunofluorescence
We recently incorporated this antibody into our immunofluorescence assays, and it's been a great addition. The antibody consistently provides clear and specific staining, enhancing the accuracy of our experiments. It's a highly dependable product that we now rely on for consistent results.
Megan Johnson25.Aug,21
High-Quality Reproducible Results
The antibody has proven to be incredibly reliable in our immunofluorescence applications. The results are always high-quality and reproducible, with strong affinity for IAV-HA. It has become a staple in our lab for various experiments, providing us with the confidence to proceed with complex assays. The robust performance of this antibody has significantly enhanced the quality of our data.
P**l12.Oct,23
Excellent for ELISA
When it comes to influenza diagnosis research, the Human Anti-IAV HA Antibody (FI6V3) has proven to be an invaluable asset. Using it in ELISA tests with nasal swab extracts from flu - suspected individuals, I incubated at 37°C for 60 minutes. The antibody's high sensitivity enabled detection of minute IAV HA levels, generating consistent results that significantly advanced my research.
Q&As
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Is the anti-IAV HA antibody suitable for detecting influenza A virus in cell cultures?
A: Yes, the anti-IAV HA antibody (PABL-214) is suitable for detecting influenza A virus in cell cultures. It provides specific binding to the HA protein, enabling accurate detection in infected cells.
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What is the optimal dilution for using the anti-IAV HA antibody in immunofluorescence assays?
A: The optimal dilution for using the anti-IAV HA antibody (PABL-214) in immunofluorescence assays is typically 1:100 to 1:500. It is recommended to perform a dilution series to determine the best working concentration for specific experimental conditions.
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Does the anti-IAV HA antibody cross-react with other influenza strains?
A: The anti-IAV HA antibody (PABL-214) is designed to be specific for the HA protein of influenza A virus. While it primarily targets this protein, some cross-reactivity with other strains of influenza may occur depending on the similarity of the HA protein among different strains.
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Can the anti-IAV HA antibody be used in ELISA applications?
A: Yes, the anti-IAV HA antibody (PABL-214) can be used in ELISA applications. It has been validated for use in such assays and provides reliable detection of the HA protein.
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What are the recommended storage conditions for the anti-IAV HA antibody?
A: The recommended storage condition for the anti-IAV HA antibody (PABL-214) is at -20°C or lower. For short-term storage, it can be kept at 2-8°C. To ensure stability, avoid repeated freeze-thaw cycles.
View the frequently asked questions answered by Creative Biolabs Support.
Citations
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Park, Jae-Keun, et al. "Pre-existing immunity to influenza virus hemagglutinin stalk might drive selection for antibody-escape mutant viruses in a human challenge model." Nature medicine 26.8 (2020): 1240-1246. https://doi.org/10.1038/s41591-020-0937-xThis study investigates the selection of an HA stalk antibody-escape mutant virus in human influenza challenge participants. The researchers focused on a well-characterized influenza A virus stock containing a mixed population of wild-type and mutant viruses with a nonsynonymous single-nucleotide polymorphism (SNP) in the HA stalk (A388V). They analyzed pre-existing anti-HA stalk antibody levels and post-challenge viral shedding in participants, revealing that higher pre-challenge anti-HA stalk antibody titers were associated with the selection of the mutant virus. The study found that the A388V mutation conferred resistance to some broadly neutralizing monoclonal antibodies (bNAbs) and was rapidly selected under immune pressure, highlighting a potential challenge for HA-stalk-targeting universal influenza vaccines.
Creative Biolabs provided several key reagents for this research, including the broadly neutralizing monoclonal antibodies CR9114 (Cat#: PABX-119), FI6V3 (Cat#: PABL-214), and CT149 (Cat#: PABL-213). These antibodies were essential for evaluating the binding and neutralization characteristics of the mutant and wild-type viruses. The specific contributions of these antibodies were crucial for understanding the structural changes and resistance mechanisms conferred by the A388V mutation, thereby aiding in the comprehensive analysis of HA stalk antibody escape and its implications for vaccine development.
Cite This Product
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# PABL-214, RRID: AB_3111639)
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Downloadable Resources
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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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|---|---|---|---|
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| PSBL-205 | Recombinant Human Anti-IAV HA Antibody scFv Fragment (1957) | WB, Neut, FuncS | scFv |
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| PABL-482 | Recombinant Human Anti-H1N1 HA Antibody (PABL-482) | FuncS, Neut, WB | IgG |
| PABL-483 | Recombinant Human Anti-H1N1 HA Antibody (CH67) | IF, FC | IgG |
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| PABL-485 | Recombinant Mouse Anti-H1N1 HA Antibody | WB, ELISA, FuncS | IgG |
| PABL-487 | Recombinant Human Anti-H2N2 HA Antibody (2G1 ) | FuncS | IgG |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| PFBL-479 | Recombinant Human Anti-H1N1 HA Antibody Fab Fragment (CH65) | WB, ELISA, FuncS | Fab |
| PFBL-480 | Recombinant Human Anti-H1N1 HA Antibody Fab Fragment (CH67) | Neut | Fab |
| PFBL-481 | Recombinant Human Anti-H1N1 HA Antibody Fab Fragment (PFBL-481) | Neut | Fab |
| PFBL-482 | Recombinant Mouse Anti-H1N1 HA Antibody Fab Fragment | WB, ELISA, FuncS | Fab |
| PFBL-484 | Recombinant Human Anti-H2N2 HA Antibody Fab Fragment (2G1 ) | Neut | Fab |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MRO-241CT | Anti-IAV HA Recombinant Antibody (D1-1) | WB, FC, IP, Inhib | Human antibody |
| MRO-242CT | Anti-IAV HA Recombinant Antibody (D1-2) | WB, FC, IP, Inhib | Human antibody |
| MRO-243CT | Anti-IAV HA Recombinant Antibody (D1-3) | WB, FC, IP, Inhib | Human antibody |
| MRO-244CT | Anti-IAV HA Recombinant Antibody (D1-4) | WB, FC, IP, Inhib | Human antibody |
| MRO-245CT | Anti-IAV HA Recombinant Antibody (D1-5) | WB, FC, IP, Inhib | Human antibody |
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| MRO-301CT | Anti-IAV HA Recombinant Antibody (3741 ) | WB, ELISA |
| CAT | Product Name | Application | Type |
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| MRO-282CT-S(P) | Anti-IAV HA Recombinant Antibody scFv Fragment (49A12) | WB, ELISA, IHC, Neut | |
| MRO-283CT-S(P) | Anti-IAV HA Recombinant Antibody scFv Fragment (21D12) | WB, ELISA, IHC, Neut | |
| MRO-284CT-S(P) | Anti-IAV HA Recombinant Antibody scFv Fragment (Clone 81) | WB, ELISA | |
| MRO-285CT-S(P) | Anti-IAV HA Recombinant Antibody scFv Fragment (Clone 11) | WB, ELISA | |
| MRO-286CT-S(P) | Anti-IAV HA Recombinant Antibody scFv Fragment (Clone 2) | WB, ELISA |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| PABX-070-S (P) | Recombinant Mouse Anti-HA1 Antibody scFv Fragment (Fab17/9) | WB, ELISA, Neut, FuncS | scFv |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MOR-4209 | Hi-Affi™ Recombinant Rabbit Anti-HA Monoclonal Antibody (SI287DS) | ELISA, ICC, IF, IF, FC | IgG |
| MOR-4210 | Hi-Affi™ Recombinant Rabbit Anti-HA Monoclonal Antibody (SI288DS) | ELISA, ICC, IF, IF, FC | IgG |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| HPAB-1169-FY | Llama Anti-HA Recombinant Single Domain Antibody (HPAB-1169-FY) | Neut | Llama VHH |
| HPAB-1170-FY | Llama Anti-HA Recombinant Single Domain Antibody (HPAB-1170-FY) | Neut | Llama VHH |
| HPAB-1171-FY | Llama Anti-HA Recombinant Single Domain Antibody (HPAB-1171-FY) | Neut | Llama VHH |
| HPAB-1172-FY | Llama Anti-HA Recombinant Single Domain Antibody (HPAB-1172-FY) | Neut | Llama VHH |
| HPAB-1173-FY | Llama Anti-HA Recombinant Single Domain Antibody (HPAB-1173-FY) | Neut | Llama VHH |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MHC-YF096 | DRB1*01:01/Influenza A HA (PKYVKQNTLKLAT) MHC Monomer | MHC Multimer | |
| MHC-YF098 | DRB1*04:01/Influenza A HA (PKYVKQNTLKLAT) MHC Monomer | MHC Multimer | |
| MHC-YF101 | DRB1*04:02/Influenza A HA (PKYVKQNTLKLAT) MHC Monomer | MHC Multimer | |
| MHC-YF102 | DRB1*04:05/Influenza A HA (PKYVKQNTLKLAT) MHC Monomer | MHC Multimer | |
| MHC-YF103 | DRB1*07:01/Influenza A HA (PKYVKQNTLKLAT) MHC Monomer | MHC Multimer |
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