Hi-Affi™ Rabbit Anti-APP Recombinant Antibody (clone M204)
CAT#: PABX-014
Anti-APP antibody is a Rabbit antibody of IgG class that binds to an APP. This antibody recognizes a generic epitope that is common to prefibrillar amyloid oligomers formed from many different amyloidogenic sequences.
Published Data
Gene Expression
ELISA
Figure 1 ELISA showing immunoreactivity of monoclonal M204 antibody with chromatographed S75 fractions containing tau-K18 oligomer (fractions 44-49, colored red), but not tau monomer (fractions 58-61, colored black).
A volume of 20 µl of the indicated SEC fraction was diluted into 100 µl of coating buffer and applied to ELISA plate. In fractions 46 and 61 (the highest peaks), 20 µl = 100 ng of protein/well.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
ELISA
Figure 2 ELISA showing immunoreactivity of monoclonal M204 with chromatographed S200 fractions containing tau40 oligomer (fractions 10-15, colored red) but not tau monomer (fractions 17-19, colored black).
A volume of 50 µl of the indicated SEC fraction containing tau40 oligomer (fractions 8-15) were diluted into 100 µl of coating buffer. For tau40 monomer fractions 17 and 19, 10 µl was used to coat ELISA plates rather than 50 µl because the peak monomer fraction was about 5 times the intensity of the oligomer peak. For reference, the amount of protein used to coat the ELISA plate in fractions 13 and 19 (the highest peaks) was 100 ng of protein.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Figure 3 Epitope mapping using a peptide array with overlapping synthetic tau peptides shows M204 binding most strongly to peptides containing the sequences KVQIINK and SVQIVY (in red).
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Figure 4 Seeding by IL15-mediated tau-K18 purified oligomers in HEK293 biosensor cells expressing YFP-tagged tau-K18. C, as in B, except following treatment of tau-K18 oligomer with 10 μm M204 antibody.
Seeding inhibition by M204 is interpreted based on the appearance of fewer puncta compared with B. Representative cells containing aggregated tau are marked by red arrows, and cells without by white arrows.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Figure 5 Quantification of seeding from representative images shown in B and C.
Tau oligomer seeding and inhibition by M204 antibody were determined by calculating the number of normalized puncta per well of a 96-well-plate. Statistical analysis was performed using one-way ANOVA (****, p < 0.0001; ***, p < 0.0006; **, p < 0.003) and Tukey's multiple comparison in GraphPad Prism. Error bars show the S.D. of three replicates.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
WB
Figure 6 Western blotting of tissue from the brain of a donor with AD, fractionated into: crude brain homogenate, Sarkosyl-insoluble, and Sarkosyl-soluble fractions, each probed with M204 or A11 anti-oligomer antibodies.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
ELISA
Figure 7 M204-scFv antibody delays in vitro aggregation.
B-D, ELISA showing immunoreactivity of the corresponding tau-K18 oligomer fractions from A with M204-scFv (B) monomer, (C) dimer, and (D) trimer. E-G, M204-scFv mediated inhibition of tau-K18 aggregation. Aggregation was measured by ThT fluorescence with M204-scFv added at molar ratios of 1:1 and 1:0.5 (tau-K18: M204-scFv), as indicated. Inhibition of IL15-induced tau aggregation by M204-scFv is shown for the M204-scFv monomer (E), dimer (F), and trimer (G). Tau-k18 is a positive control that was run in parallel but without the addition of M204-scFv, and is the same in aggregation experiments shown in E-G, colored black.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Figure 8 B-E, quantification of the inhibitory effect of M204-scFv antibodies on inhibition of seeding by Sarkosyl-insoluble fractions from AD brain tissue, measured in HEK293 biosensor cells expressing YFP-tagged tau-K18.
Statistical analysis was performed using one-way ANOVA (****, p < 0.0001; ***, p < 0.0002; **, p < 0.001; *, p < 0.01, n.s., no significance) followed by a Tukey's multiple comparison test in GraphPad Prism. Error bars show the S.D. of three replicates.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Inhib
Figure 9 Representative images of seeding and inhibition in HEK293 biosensor cells expressing YFP-tagged tau-K18.
Cells seeded with the Sarkosyl-insoluble fraction from AD Donor 1 without pre-treatment with M204-scFv (left panel) or no inhibitor (No I), and following overnight incubation with M204-scFv trimer, dimer, or monomer (as indicated). The seeding can vary dramatically from different regions of a given brain tissue section, and this reflects a naturally occurring nonuniform distribution of tau pathology in the brain. Representative cells that contain aggregates are marked by red arrows, and cells without by white arrows.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
Inhib
Figure 10 A-D, tests of the effectiveness of M204-scFv for inhibition of tau fibril seeding by diseased brain extracts in our HEK293 biosensor cell-based assay.
M204-scFv dimer and trimer inhibit seeding by extract from (A) CTE Donor 1, but not (B) CTE Donor 2, (C) CBD donor, or (D) a donor with a P301L familial mutation. Statistical analysis was performed using one-way ANOVA (****, p < 0.0001; ***, p < 0.0008; **, p < 0.005; *, p < 0.01, n.s., no significance) followed by a Tukey's multiple comparison test in GraphPad Prism. Error bars show the S.D. of three replicates.
Abskharon, R., Seidler, P. M., Sawaya, M. R., Cascio, D., Yang, T. P., Philipp, S., ... & Eisenberg, D. S. (2020). Crystal structure of a conformational antibody that binds tau oligomers and inhibits pathological seeding by extracts from donors with Alzheimer's disease. Journal of Biological Chemistry, 295(31), 10662-10676.
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Subcellular Location
Figure 1 IF staining of human cell line U-2 OS
Immunofluorescent staining of human cell line U-2 OS shows localization to the Golgi apparatus & vesicles.
* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/1462/if_selected.jpg
Subcellular Location
Figure 2 IF staining of human cell line A-431
Immunofluorescent staining of human cell line A-431 shows localization to the Golgi apparatus.
* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/1462/60_E2_1_blue_red_green.jpg
Subcellular Location
Figure 3 IF staining of human cell line U-251 MG
Immunofluorescent staining of human cell line U-251 MG shows localization to the Golgi apparatus.
* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/1462/61_E2_1_blue_red_green.jpg
Normal Tissue
Figure 4 Cerebral cortex
Neuronal cells
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/images/157/2073_B_7_5.jpg
RNA Expression
Figure 5 RNA cell line category: Low cell line specificity
Cell lines ordered by descending RNA expression order.
* Image credit: Image credit: Human Protein Atlas https://v21.proteinatlas.org/ENSG00000142192-APP
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Specifications
- Immunogen
- The details of the immunogen for this antibody are not available.
- Host Species
- Rabbit
- Derivation
- Rabbit
- Type
- Rabbit IgG
- Specificity
- APP
- Species Reactivity
- Human
- Clone
- M204
- Applications
- WB, ELISA, FuncS
Product Property
- Purity
- >95% as determined by analysis by SDS-PAGE
- Concentration
- Please refer to the vial label for the specific concentration.
- Buffer
- PBS
- Preservative
- No preservatives
- Storage
- Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.
Target
- Alternative Names
- APP; amyloid beta (A4) precursor protein; AAA; AD1; PN2; ABPP; APPI; CVAP; ABETA; PN-II; CTFgamma; amyloid beta A4 protein; preA4; protease nexin-II; peptidase nexin-II; beta-amyloid peptide; beta-amyloid peptide (1-40); beta-amyloid peptide (1-42); alzheimer disease amyloid protein; cerebral vascular amyloid peptide
- Gene ID
- 351
- UniProt ID
- P05067
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Clear Datasheets Available
The datasheets provided are clear and detailed, featuring comprehensive information about the antibody's characteristics and usage. This transparency is greatly appreciated.
Continual Excellence
The antibody was easy to use, even for inexperienced team members, thanks to the detailed instructions that came with it.
Sturdy Performance
The comprehensive datasheet that came with the antibody was really beneficial; it included all the characteristics you needed as well as usage advice.
Q&As
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How should the working dilution be calculated when using this antibody in various assays?
A: The best functioning dilutions ought to be ascertained by experimentation. The datasheet lists typical starting dilutions; however, depending on the particular experiment circumstances, modifications can be required.
-
How can the effectiveness of this antibody in an experiment be verified?
A: Testing it on known positive and negative samples, applying the proper controls, and contrasting the outcomes with predetermined benchmarks are all part of the validation process.
-
Does this antibody require any particular handling?
A: When preparing reagents for use in research, always utilize sterile techniques and handle the antibody with care to prevent contamination.
View the frequently asked questions answered by Creative Biolabs Support.
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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.
Datasheet
MSDS
COA
Certificate of Analysis LookupTo download a Certificate of Analysis, please enter a lot number in the search box below. Note: Certificate of Analysis not available for kit components.
Lot Number:
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- CAT
- Product Name
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| TAB-0789CLV-F(E) | Human Anti-APP Recombinant Antibody; Fab Fragment (TAB-0789CLV-F(E)) | ELISA | Chimeric (mouse/human) Fab |
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| TAB-0789CLV-S(P) | Mouse Anti-APP Recombinant Antibody; scFv Fragment (TAB-0789CLV-S(P)) | ELISA | Mouse scFv |
| TAB-0802CLV-S(P) | Mouse Anti-APP Recombinant Antibody; scFv Fragment (TAB-0802CLV-S(P)) | ELISA | Mouse scFv |
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| CAT | Product Name | Application | Type |
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| PABX-014-F (E) | Hi-Affi™ Rabbit Anti-APP Recombinant Antibody (clone M204); Fab Fragment | WB, ELISA, FuncS | Rabbit Fab |
| MOR-0220 | Hi-Affi™ Rabbit Anti-APP Recombinant Antibody (clone DS220AB) | ELISA, WB | Rabbit IgG |
| MOR-4539 | Hi-Affi™ Rabbit Anti-APP Recombinant Antibody (clone TH47DS) | IHC-P, ELISA | Rabbit IgG |
| MOR-0082-FY | Rabbit Anti-APP Recombinant Antibody (clone AFY0053) | ICC, IHC-P, WB | Rabbit IgG |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| EPAF-0566CQ | Mouse Anti-APP Recombinant Antibody (clone 6E10) | ELISA | Mouse IgG |
| EPAF-0567CQ | Mouse Anti-APP Recombinant Antibody (clone 4G8) | ELISA, IHC | Mouse IgG2b |
| EPAF-0568CQ | Human Anti-APP Recombinant Antibody (clone HyL5) | ELISA, Inhib | Human IgM |
| EPAF-0569CQ | Mouse Anti-APP Recombinant Antibody (clone AB1-42) | IHC | Mouse IgG |
| EPAF-0570CQ | Human Anti-APP Recombinant Antibody (clone L11.3) | IHC, ELISA, Inhib | Human IgM |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| NS-001CN | Human Anti-APP Recombinant Antibody (clone AS) | WB | Human IgG |
| NS-007CN | Mouse Anti-APP Recombinant Antibody (NS-007CN) | ELISA, FuncS | Mouse IgG1 |
| NS-001CN-F(E) | Human Anti-APP Recombinant Antibody (clone AS); Fab Fragment | WB | Human Fab |
| NS-007CN-F(E) | Mouse Anti-APP Recombinant Antibody; Fab Fragment (NS-007CN-F(E)) | ELISA | Mouse Fab |
| NS-001CN-S(P) | Human Anti-APP Recombinant Antibody (clone AS); scFv Fragment | WB | Human scFv |
| CAT | Product Name | Application | Type |
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| AFC-TAB-228 | Afuco™ Anti-APP ADCC Recombinant Antibody, ADCC Enhanced (AFC-TAB-228) | FuncS, IF, Neut, ELISA, FC | ADCC enhanced antibody |
| AFC-TAB-229 | Afuco™ Anti-APP ADCC Recombinant Antibody, ADCC Enhanced (AFC-TAB-229) | FuncS, IF, Neut, ELISA, FC, IP | ADCC enhanced antibody |
| AFC-TAB-227 | Afuco™ Anti-APP ADCC Recombinant Antibody, ADCC Enhanced (AFC-TAB-227) | ELISA, IP, FC, FuncS, Neut | ADCC enhanced antibody |
| AFC-TAB-226 | Afuco™ Anti-APP ADCC Recombinant Antibody, ADCC Enhanced (AFC-TAB-226) | IF, IP, Neut, FuncS, ELISA | ADCC enhanced antibody |
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| HPAB-0008-YJ | Mouse Anti-APP Recombinant Antibody (clone 508F) | ELISA | Mouse IgM |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| HPAB-0525-WJ-F(E) | Human Anti-APP Recombinant Antibody (clone h12A11); Fab Fragment | ELISA | Humanized Fab |
| HPAB-0970-FY-S(P) | Mouse Anti-APP Recombinant Antibody; scFv Fragment (HPAB-0970-FY-S(P)) | ELISA | Mouse scFv |
| HPAB-0971-FY-S(P) | Mouse Anti-APP Recombinant Antibody; scFv Fragment (HPAB-0971-FY-S(P)) | ELISA | Mouse scFv |
| HPAB-0973-FY-S(P) | Human Anti-APP Recombinant Antibody (clone E12); Fab Fragment | ELISA | Humanized Fab |
| HPAB-0974-FY-S(P) | Human Anti-APP Recombinant Antibody (clone 1C10); Fab Fragment | ELISA | Humanized Fab |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| HPAB-0525-WJ-S(P) | Human Anti-APP Recombinant Antibody (clone h12A11); scFv Fragment | ELISA | Humanized scFv |
| HPAB-0970-FY-F(E) | Mouse Anti-APP Recombinant Antibody; Fab Fragment (HPAB-0970-FY-F(E)) | ELISA | Mouse Fab |
| HPAB-0971-FY-F(E) | Mouse Anti-APP Recombinant Antibody; Fab Fragment (HPAB-0971-FY-F(E)) | ELISA | Mouse Fab |
| HPAB-0973-FY-F(E) | Human Anti-APP Recombinant Antibody (clone E12); scFv Fragment | ELISA | Humanized scFv |
| HPAB-0974-FY-F(E) | Human Anti-APP Recombinant Antibody (clone 1C10); scFv Fragment | ELISA | Humanized scFv |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-0424-XY17 | AbPlus™ Anti-APP Magnetic Beads (DE2B4) | IP, Protein Purification |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-1024-XY23 | Mouse Anti-NHP APP Recombinant Antibody (VS-1024-XY23) | WB, ICC | Mouse IgG1 |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-0325-FY6 | Human Anti-APP (clone 2A7) scFv-Fc Chimera | ELISA | Human IgG1, scFv-Fc |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-0425-YC204 | Recombinant Anti-APP Vesicular Antibody, EV Displayed (VS-0425-YC204) | ELISA, FC, Neut, Cell-uptake |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-0525-XY469 | Anti-APP Immunohistochemistry Kit | IHC | |
| VS-0525-XY470 | Anti-Mouse APP Immunohistochemistry Kit | IHC | |
| VS-0525-XY471 | Anti-Monkey APP Immunohistochemistry Kit | IHC |
| CAT | Product Name | Application | Type |
|---|---|---|---|
| VS-0525-YC2 | Recombinant Anti-APP Biparatopic Antibody, Tandem sdAb | ELISA, Inhib | Tandem sdAb |
| VS-0525-YC18 | Recombinant Anti-APP (AA 3-7 x AA 12-28) Biparatopic Antibody, Tandem scFv (Clone h12A11v1 x Solanezumab) | ELISA, Neut, FC, IF, IP, WB | Tandem scFv |
| VS-0525-YC196 | Recombinant Anti-APP (AA 1-5 x AA 13-28) Biparatopic Antibody, Tandem scFv | ELISA, FC, IF, IP, Neut, WB | Tandem scFv |
| VS-0525-YC198 | Recombinant Anti-APP (AA 13-28 x AA 33-40) Biparatopic Antibody, Tandem scFv (Solanezumab x Ponezumab) | ELISA, FC, IF, IP, Neut, WB | Tandem scFv |
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