Rabbit HP1 alpha/CBX5 Polyclonal Antibody | anti-CBX5 antibody
Anti-HP1 alpha/CBX5 Antibody
IHC-P: 0.5-1ug/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|
Western Blot (WB)
(Figure 1. Western blot analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).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 U-87MG whole cell lysatesLane 2: human K562 whole cell lysatesLane 3: human Jurkat whole cell lysatesLane 4: human Raji whole cell lysatesLane 5: rat PC-12 whole cell lysatesLane 6: 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-HP1 alpha/CBX5 antigen affinity purified polyclonal antibody (Catalog # MBS1753550) 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 1:10000 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 HP1 alpha/CBX5 at approximately 22KD. The expected band size for HP1 alpha/CBX5 is at 22KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 7. IF analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (MBS1753550).HP1 alpha/CBX5 was detected in immunocytochemical section of Hela 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 2μg/mL rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550) 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 8. Flow Cytometry analysis of K562 cells using anti-HP1 alpha/CBX5 antibody (MBS1753550).Overlay histogram showing K562 cells stained with MBS1753550 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HP1 alpha/CBX5 Antibody (MBS1753550,1μg/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. Bannister, A. J., Zegerman, P., Partridge, J. F., Miska, E. A., Thomas, J. O., Allshire, R. C., Kouzarides, T. Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain. Nature 410: 120-124, 2001.
3. Cheutin, T., McNairn, A. J., Jenuwein, T., Gilbert, D. M., Singh, P. B., Misteli, T. Maintenance of stable heterochromatin domains by dynamic HP1 binding. Science 299: 721-725, 2003.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a highly conserved nonhistone protein, which is a member of the heterochromatin protein family. The protein is enriched in the heterochromatin and associated with centromeres. The protein has a single N-terminal chromodomain which can bind to histone proteins via methylated lysine residues, and a C-terminal chromo shadow-domain (CSD) which is responsible for the homodimerization and interaction with a number of chromatin-associated nonhistone proteins. The encoded product is involved in the formation of functional kinetochore through interaction with essential kinetochore proteins. The gene has a pseudogene located on chromosome 3. Multiple alternatively spliced variants, encoding the same protein, have been identified. [provided by RefSeq, Jul 2008]
Uniprot Description
HP1 alpha: Component of heterochromatin that recognizes and binds histone H3 tails methylated at 'Lys-9' (H3K9me), leading to epigenetic repression. In contrast, it is excluded from chromatin when 'Tyr-41' of histone H3 is phosphorylated (H3Y41ph). Can interact with lamin-B receptor (LBR). This interaction can contribute to the association of the heterochromatin with the inner nuclear membrane. Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins. Interacts with SUV420H1 and SUV420H2. Interacts with HP1BP3. Interacts directly with ATRX, CHAF1A, LBR, NIPBL, SP100, STAM2 and TRIM28 via the chromoshadow domain. Can interact directly with CBX3 via the chromoshadow domain. Interacts with histone H3 methylated at 'Lys- 9'. Interacts with BAHD1, MIS12 and DSN1. Interacts with POGZ; POGZ and PXVXL motif-containing proteins such as INCENP and TRIM28 compete for interaction with CBX5. Interacts with INCENP and TRIM24. Interacts with JC virus agnoprotein; this interaction induces the dissociation of CBX5 from LBR, resulting in destabilization of the nuclear envelope. Interacts with CHAMP1. Interacts with ASXL1.
Protein type: Transcription, coactivator/corepressor
Chromosomal Location of Human Ortholog: 12q13.13
Cellular Component: kinetochore; nucleoplasm; PML body; histone methyltransferase complex; transcriptional repressor complex; histone deacetylase complex; nucleolus; nuclear envelope; nuclear heterochromatin; nucleus; chromocenter
Molecular Function: protein binding, bridging; protein binding; protein homodimerization activity; histone deacetylase binding; chromatin binding; methylated histone residue binding
Biological Process: viral reproduction; negative regulation of transcription, DNA-dependent; blood coagulation
Research Articles on CBX5
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Product Notes
The CBX5 cbx5 (Catalog #AAA1753550) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-HP1 alpha/CBX5 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's HP1 alpha/CBX5 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Mouse, Rat| IHC-P: 0.5-1ug/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 CBX5 cbx5 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 "HP1 alpha/CBX5, 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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