Western Blot (WB)
(Figure 1. Western blot analysis of WAC using anti-WAC antibody (MBS1753680).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 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human U20S whole cell lysatesLane 6: human SK-OV-3 whole cell lysatesLane 7: rat brain tissue lysatesLane 8: rat PC-12 whole cell lysatesLane 9: mouse thymus tissue lysatesLane 10: mouse SP2/0 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-WAC antigen affinity purified polyclonal antibody (Catalog # MBS1753680) at 0. 25 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 1:10000 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 WAC at approximately 85KD. The expected band size for WAC is at 85KD. )
Rabbit WAC Polyclonal Antibody | anti-WAC antibody
Anti-WAC Antibody
ICC/IF: 4ug/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 WAC using anti-WAC antibody (MBS1753680).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 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human U20S whole cell lysatesLane 6: human SK-OV-3 whole cell lysatesLane 7: rat brain tissue lysatesLane 8: rat PC-12 whole cell lysatesLane 9: mouse thymus tissue lysatesLane 10: mouse SP2/0 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-WAC antigen affinity purified polyclonal antibody (Catalog # MBS1753680) at 0. 25 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 1:10000 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 WAC at approximately 85KD. The expected band size for WAC is at 85KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of WAC using anti-WAC antibody (MBS1753680).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 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human U20S whole cell lysatesLane 6: human SK-OV-3 whole cell lysatesLane 7: rat brain tissue lysatesLane 8: rat PC-12 whole cell lysatesLane 9: mouse thymus tissue lysatesLane 10: mouse SP2/0 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-WAC antigen affinity purified polyclonal antibody (Catalog # MBS1753680) at 0. 25 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 1:10000 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 WAC at approximately 85KD. The expected band size for WAC is at 85KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of WAC using anti-WAC antibody (MBS1753680).WAC was detected in immunocytochemical section of A431 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 4μg/mL rabbit anti-WAC Antibody (MBS1753680) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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 WAC using anti-WAC antibody (MBS1753680).WAC was detected in immunocytochemical section of A431 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 4μg/mL rabbit anti-WAC Antibody (MBS1753680) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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 HepG2 cells using anti-WAC antibody (MBS1753680).Overlay histogram showing HepG2 cells stained with MBS1753680 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-WAC Antibody (MBS1753680,1μg/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 HepG2 cells using anti-WAC antibody (MBS1753680).Overlay histogram showing HepG2 cells stained with MBS1753680 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-WAC Antibody (MBS1753680,1μg/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. Nagase, T., Nakayama, M., Nakajima, D., Kikuno, R., Ohara, O. Prediction of the coding sequences of unidentified human genes. XX. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 8: 85-95, 2001.
3. Xu, G. M., Arnaout, M. A. WAC, a novel WW domain-containing adapter with a coiled-coil region, is colocalized with splicing factor SC35. Genomics 79: 87-94, 2002.
NCBI and Uniprot Product Information
Observed: 75 kDa
NCBI Description
The protein encoded by this gene contains a WW domain, which is a protein module found in a wide range of signaling proteins. This domain mediates protein-protein interactions and binds proteins containing short linear peptide motifs that are proline-rich or contain at least one proline. This gene product shares 94% sequence identity with the WAC protein in mouse, however, its exact function is not known. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Dec 2008]
Uniprot Description
WAC: Acts as a linker between gene transcription and histone H2B monoubiquitination at 'Lys-120' (H2BK120ub1). Interacts with the RNA polymerase II transcriptional machinery via its WW domain and with RNF20-RNF40 via its coiled coil region, thereby linking and regulating H2BK120ub1 and gene transcription. Regulates the cell-cycle checkpoint activation in response to DNA damage. Positive regulator of amino acid starvation-induced autophagy. May negatively regulate the ubiquitin proteasome pathway. 4 isoforms of the human protein are produced by alternative splicing.
Protein type: Adaptor/scaffold
Chromosomal Location of Human Ortholog: 10p12.1|10p12.1-p11.2
Cellular Component: nucleoplasm; spliceosome; nuclear speck; nucleus
Molecular Function: protein binding; chromatin binding
Biological Process: histone monoubiquitination; transcription, DNA-dependent; positive regulation of transcription, DNA-dependent; negative regulation of proteasomal ubiquitin-dependent protein catabolic process; response to DNA damage stimulus; positive regulation of macroautophagy
Research Articles on WAC
Similar Products
Product Notes
The WAC wac (Catalog #AAA1753680) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-WAC Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's WAC 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: 0.1-0.25ug/ml|Human, Mouse, Rat| ICC/IF: 4ug/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 WAC wac 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 "WAC, 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
If you are ready to order, navigate to Shopping Cart and get ready to checkout.
