Recombinant Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19803] to HP1 gamma/CBX3
- Suitable for: WB, Flow Cyt (Intra), IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HP1 gamma/CBX3 antibody [EPR19803]
See all HP1 gamma/CBX3 primary antibodies -
Description
Rabbit monoclonal [EPR19803] to HP1 gamma/CBX3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra), IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal brain and fetal kidney lysates; HepG2, HeLa, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and spleen lysates. IHC-P: Human colon and cervix carcinoma tissues; Mouse testis tissue. ICC/IF: NIH/3T3 and HeLa cells. Flow Cyt (intra): HeLa and NIH/3T3 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19803 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213167 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (1) |
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
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Flow Cyt (Intra) |
1/60.
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IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This application is recommended for human and mouse only. |
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ICC/IF | (1) |
1/100.
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Notes |
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WB
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa). |
Flow Cyt (Intra)
1/60. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This application is recommended for human and mouse only. |
ICC/IF
1/100. |
Target
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Function
Seems to be involved in transcriptional silencing in heterochromatin-like complexes. Recognizes and binds histone H3 tails methylated at 'Lys-9', leading to epigenetic repression. May contribute to the association of the heterochromatin with the inner nuclear membrane through its interaction with lamin B receptor (LBR). Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins. -
Sequence similarities
Contains 2 chromo domains. -
Post-translational
modificationsPhosphorylated by PIM1. Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis. -
Cellular localization
Nucleus. Associates with euchromatin and is largely excluded from constitutive heterochromatin. May be associated with microtubules and mitotic poles during mitosis. - Information by UniProt
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Database links
- Entrez Gene: 11335 Human
- Entrez Gene: 12417 Mouse
- Entrez Gene: 297093 Rat
- Omim: 604477 Human
- SwissProt: Q13185 Human
- SwissProt: P23198 Mouse
- Unigene: 381189 Human
- Unigene: 280968 Mouse
see all -
Alternative names
- CBX 3 antibody
- CBX3 antibody
- CBX3_HUMAN antibody
see all
Images
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Lanes:
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: CBX3 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HUVEC whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab213167 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab213167 was shown to recognize HP1 gamma/CBX3 when HP1 gamma/CBX3 knockout samples were used, along with additional cross-reactive bands. Wild-type and HP1 gamma/CBX3 knockout samples were subjected to SDS-PAGE. Ab213167 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDa
Exposure time: 3 minutes -
All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2: 15 seconds.
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All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat spleen lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 21 kDa
Observed band size: 21 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 30 seconds; Lane 5: 10 seconds; Lane 6,7,8 and 9: 30 seconds.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on epithelial cells of Human colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on cancer cells of Human cervix carcinoma is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on Mouse testis is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HP1 gamma/CBX3 with ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on NIH/3T3 cells.
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab213167 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HP1 gamma/CBX3 with ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HeLa cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab213167 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HP1 gamma/CBX3with ab213167 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HP1 gamma/CBX3 with ab213167 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal- Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab213167 has been referenced in 3 publications.
- Sun X et al. Autism-associated protein POGZ controls ESCs and ESC neural induction by association with esBAF. Mol Autism 13:24 (2022). PubMed: 35650610
- Jeon S et al. Ablation of CRBN induces loss of type I collagen and SCH in mouse skin by fibroblast senescence via the p38 MAPK pathway. Aging (Albany NY) 13:6406-6419 (2021). PubMed: 33658395
- Zhou W et al. lncRNA KCNQ1OT1 reverses the effect of sevoflurane on hepatocellular carcinoma progression via regulating the miR-29a-3p/CBX3 axis. Braz J Med Biol Res 54:e10213 (2021). PubMed: 34008749