Western Blot (WB)
(Figure 1. Western blot analysis of HAP1 using anti-HAP1 antibody (MBS1753449).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: mouse Neuro-2a whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-HAP1 antigen affinity purified polyclonal antibody (Catalog # MBS1753449) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for HAP1 at approximately 80KD. The expected band size for HAP1 is at 80KD. )
Rabbit HAP1 Polyclonal Antibody | anti-HAP1 antibody
Anti-HAP1 Antibody
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of HAP1 using anti-HAP1 antibody (MBS1753449).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: mouse Neuro-2a whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-HAP1 antigen affinity purified polyclonal antibody (Catalog # MBS1753449) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for HAP1 at approximately 80KD. The expected band size for HAP1 is at 80KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of HAP1 using anti-HAP1 antibody (MBS1753449).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: mouse Neuro-2a whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-HAP1 antigen affinity purified polyclonal antibody (Catalog # MBS1753449) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for HAP1 at approximately 80KD. The expected band size for HAP1 is at 80KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of HAP1 using anti- HAP1 antibody (MBS1753449).HAP1 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- HAP1 Antibody (MBS1753449) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of HAP1 using anti- HAP1 antibody (MBS1753449).HAP1 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- HAP1 Antibody (MBS1753449) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of U937 cells using anti-HAP1 antibody (MBS1753449).Overlay histogram showing U937 cells stained with MBS1753449 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HAP1 Antibody (MBS1753449, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of U937 cells using anti-HAP1 antibody (MBS1753449).Overlay histogram showing U937 cells stained with MBS1753449 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HAP1 Antibody (MBS1753449, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Chan, E. Y. W., Nasir, J., Gutekunst, C. -A., Coleman, S., Maclean, A., Maas, A., Metzler, M., Gertsenstein, M., Ross, C. A., Nagy, A., Hayden, M. R. Targeted disruption of Huntingtin-associated protein-1 (Hap1) results in postnatal death due to depressed feeding behavior. Hum. Molec. Genet. 11: 945-959, 2002.
3. Dragatsis, I., Zeitlin, S., Dietrich, P. Huntingtin-associated protein 1 (Hap1) mutant mice bypassing the early postnatal lethality are neuroanatomically normal and fertile but display growth retardation. Hum. Molec. Genet. 13: 3115-3125, 2004.
NCBI and Uniprot Product Information
NCBI Description
Huntington's disease (HD), a neurodegenerative disorder characterized by loss of striatal neurons, is caused by an expansion of a polyglutamine tract in the HD protein huntingtin. This gene encodes a protein that interacts with huntingtin, with two cytoskeletal proteins (dynactin and pericentriolar autoantigen protein 1), and with a hepatocyte growth factor-regulated tyrosine kinase substrate. The interactions with cytoskeletal proteins and a kinase substrate suggest a role for this protein in vesicular trafficking or organelle transport. Several alternatively spliced transcript variants encoding different isoforms have been described for this gene. [provided by RefSeq, Jul 2008]
Uniprot Description
HAP1: Associates specifically with huntingtin. This binding is enhanced by an expanded polyglutamine repeat. 4 isoforms of the human protein are produced by alternative splicing.
Protein type: Motility/polarity/chemotaxis; Cytoskeletal
Chromosomal Location of Human Ortholog: 17q21.2-q21.3
Cellular Component: synaptic vesicle; cytoskeleton; mitochondrion; axon; lysosome; endoplasmic reticulum; autophagic vacuole; inclusion body; nucleus; cell junction; actin cytoskeleton
Molecular Function: protein binding; brain-derived neurotrophic factor binding
Biological Process: positive regulation of neurogenesis; positive regulation of inositol-1,4,5-triphosphate receptor activity; exocytosis; nerve growth factor receptor signaling pathway; positive regulation of synaptic transmission, GABAergic; hypothalamus cell differentiation; retrograde axon cargo transport; regulation of exocytosis; synaptic transmission; protein transport; protein localization; cell projection organization and biogenesis; positive regulation of epidermal growth factor receptor signaling pathway; vesicle transport along microtubule; positive regulation of neurotrophin production; autophagy; cerebellum development; brain development; anterograde axon cargo transport
Research Articles on HAP1
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Product Notes
The HAP1 hap1 (Catalog #AAA1753449) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-HAP1 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's HAP1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Mouse, Rat| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the HAP1 hap1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "HAP1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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