Rabbit Recombinant Monoclonal HP1 alpha antibody. Heterochromatin marker. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 16 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | ICC/IF | IP | WB | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Expected | Tested | Expected |
Rat | Tested | Expected | Expected | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/1000 | Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/10 - 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes For unpurified use at 1/10000 - 1/50000. |
Species Rat | Dilution info 1/1000 | Notes For unpurified use at 1/10000 - 1/50000. |
Species Human | Dilution info 1/1000 | Notes For unpurified use at 1/10000 - 1/50000. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/10 - 1/100 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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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) (PubMed:19783980). May contribute to the association of heterochromatin with the inner nuclear membrane by interactions with the lamin-B receptor (LBR) (PubMed:19783980). Involved in the formation of kinetochore through interaction with the MIS12 complex subunit NSL1 (PubMed:19783980, PubMed:20231385). Required for the formation of the inner centromere (PubMed:20231385).
HP1A, HP1A, CBX5, Chromobox protein homolog 5, Antigen p25, Heterochromatin protein 1 homolog alpha, HP1 alpha
Rabbit Recombinant Monoclonal HP1 alpha antibody. Heterochromatin marker. Suitable for IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 16 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR5777
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
HP1 alpha also known as HP1a or HP1 protein is a well-studied member of the heterochromatin protein 1 family. It plays a mechanical role in chromatin organization and gene regulation by binding to histone H3 when it's methylated at lysine 9 (H3K9me). This protein has a molecular mass of approximately 22-25 kDa. HP1 alpha is commonly expressed in the nuclei of eukaryotic cells where it localizes to heterochromatin regions and influences chromatin compaction.
Heterochromatin protein 1 alpha acts as a modulator of gene silencing. It is part of a larger protein complex that contains various chromatin modifiers responsible for maintaining the silent state of certain genes. By mediating these interactions HP1 alpha helps regulate transcription replication timing and DNA repair processes. This regulation is essential for the structural maintenance of heterochromatin and plays a part in chromosomal stability.
HP1 alpha is integral in the epigenetic regulation pathway along with other well-known proteins including SUV39H1 and SUV39H2 which are histone methyltransferases. HP1 alpha also links to the DNA damage response pathway where it recruits repair factors to maintain genomic integrity. Through these pathways HP1 alpha interacts closely with proteins like BRCA1 emphasizing its role in cellular homeostasis and response to DNA damage.
Researchers associate HP1 alpha with cancer and developmental disorders. Alterations in HP1 alpha expression or function can lead to disruptions in chromatin organization contributing to oncogenesis by affecting tumor suppressor genes. Furthermore HP1 alpha links to disorders like ICF syndrome through its relationship with the DNA methylation process where it interacts with proteins such as DNMT3B. Understanding these connections provides insights into therapeutic approaches targeting epigenetic modifications in disease contexts.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Purified ab109028 at 1/30 dilution (2μg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.
Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+): ab109028 + MCF7 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109028 in MCF7 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 22 kDa
All lanes: Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)
Predicted band size: 22 kDa
We are unsure how to define the extra bands.
All lanes: Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg
Lane 3: Mouse brain lysate at 20 µg
Lane 4: Rat brain lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
We are unsure how to define the extra bands.
Purified ab109028 at 1/30 dilution (2μg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: HP1 alpha knockout HAP1 cell lysate (20 μg)
Lane 3: HeLa cell lysate (20 μg)
Lane 4: MCF7 cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109028 observed at 28 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab109028 was shown to specifically react with HP1 alpha when HP1 alpha knockout samples were used. Wild-type and HP1 alpha knockout samples were subjected to SDS-PAGE. ab109028 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/20 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028)
Predicted band size: 22 kDa, 24 kDa
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HP1 alpha with Purified ab109028 at 1/30 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
ab109028 staining HP1 alpha in wild-type HAP1 cells (top panel) and HP1 alpha knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109028 at 1/250 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat pancreas tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
ab109028 staining HP1 alpha in HeLa (human cervical adenocarcinoma epithelial cell). The cells were fixed with 100% methanol, permeabilized with 0.1% TritonX-100. The cells were then incubated with ab109028 at 1:100 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1:1000 dilution (shown in green). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1:200 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Confocal image showing nuclear staining in HeLa cell line.
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