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- References [2]
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- Validations
- Western blot [1]
- Other assay [2]
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- Product number
- PA1-41610 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Humanin
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide corresponding to residues 12-24 of human humanin.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µl
- Storage
- -20°C
Submitted references Humanin Blocks the Aggregation of Amyloid-β Induced by Acetylcholinesterase, an Effect Abolished in the Presence of IGFBP-3.
Interaction of amyloid beta with humanin and acetylcholinesterase is modulated by ATP.
Price D, Dorandish S, Williams A, Iwaniec B, Stephens A, Marshall K, Guthrie J, Heyl D, Evans HG
Biochemistry 2020 Jun 2;59(21):1981-2002
Biochemistry 2020 Jun 2;59(21):1981-2002
Interaction of amyloid beta with humanin and acetylcholinesterase is modulated by ATP.
Atali S, Dorandish S, Devos J, Williams A, Price D, Taylor J, Guthrie J, Heyl D, Evans HG
FEBS open bio 2020 Dec;10(12):2805-2823
FEBS open bio 2020 Dec;10(12):2805-2823
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Humanin in recombinant fusion protein. Samples were incubated in Humanin polyclonal antibody (Product # PA1-41610). (A) recombinant fusion protein containing amino acids 12-24 and (B) fusion partner without these amino acids.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 5 Fig. ATP increases the amount of HN found in a complex with Abeta in both A549 and H1299 cell media while AChE found in a complex with Abeta is decreased by the addition of ATP to A549 cell media. Cells (0.2 x 10 5 cells per well) were seeded in 96-well plates in 10% FBS-supplemented media. The next day, the cells were incubated in serum-free medium for 72 h. Specific antibodies were added (1 : 1000 dilution) to ELISA wells. After blocking the wells, 300 muL of the A549 (A, B) or H1299 (C, D) cell-conditioned medium (0.5 mug*muL -1 ), 72 h postserum starvation, was added. The proteins/peptides were detected using their corresponding primary antibodies and then processed as described in Materials and methods section. Each column represents the mean +- SD of three independent separate experiments, each performed in triplicate. Data processing was carried out using the graphpad 8.4.3 software. Asterisks (**) indicate a statistically significant difference between each treatment relative to samples without ATP. Absence of asterisks indicates no significance, Mann-Whitney test, ** P < 0.01.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 6 Fig. Exogenously added ATP increases HN bound to Abeta from media of A549 and H1299 cells using 82E1 antibodies, resulting in decreased binding of exogenously added HN to Abeta. Conversely, less AChE is found in a complex with Abeta in the presence of added ATP in A549 cells. Anti-Abeta-specific antibodies (82E1) were added (1 : 1000 dilution) to ELISA plate wells. The wells were blocked, and 300 uL of conditioned media (0.5 ug*uL -1 ) of A549 cells (A) or H1299 cells (B), 72 h after serum starvation, was added without or with ATP and increasing HN concentrations, and then, the HN and AChE bound were detected using the corresponding specific primary antibodies and processed as described in Materials and methods. Fold change relative to controls incubated with all components except the primary antibodies was calculated and fit, using the graphpad prism 8.4.3 software, with a nonlinear regression curve. The data represent the mean +- SD of three separate experiments, each performed in triplicate.
