Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

Rabbit Recombinant Monoclonal HP1 gamma/CBX3 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples.

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Images

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	WB	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes This application is recommended for human and mouse only. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes This application is recommended for human and mouse only. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Target data

See full target information CBX3

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. Contributes to the conversion of local chromatin to a heterochromatin-like repressive state through H3 'Lys-9' trimethylation, mediates the recruitment of the methyltransferases SUV39H1 and/or SUV39H2 by the PER complex to the E-box elements of the circadian target genes such as PER2 itself or PER1. Mediates the recruitment of NIPBL to sites of DNA damage at double-strand breaks (DSBs) (PubMed:28167679).

Alternative names

Chromobox protein homolog 3, HECH, Heterochromatin protein 1 homolog gamma, Modifier 2 protein, HP1 gamma, CBX3

Recommended products

Rabbit Recombinant Monoclonal HP1 gamma/CBX3 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR19803
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab251540 is the carrier-free version of Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167.

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The HP1 gamma protein also known as CBX3 is an essential component of the chromatin organization machinery. It has a molecular weight of approximately 22-25 kDa. HP1 gamma is expressed in a variety of tissues including brain liver and muscle indicating its broad functional involvement across different biological systems. It exhibits a high affinity for binding to histone H3 when it is methylated facilitating the formation of heterochromatin which is key to regulating gene expression and maintaining genome stability.

Biological function summary

HP1 gamma plays a significant role in gene silencing and chromatin structure maintenance. It forms part of a protein complex that includes chromatin remodeling enzymes and transcriptional repressors which help regulate access to genomic DNA. HP1 gamma's association with other chromatin-associated proteins helps modulate the transcriptional response to cellular stimuli ensuring appropriate gene expression patterns needed for normal cell function and development.

Pathways

HP1 gamma is involved in heterochromatin formation and maintenance pathways working alongside other proteins like SUV39H1 and HP1 alpha. Through these pathways HP1 gamma ensures the compact organization of repetitive DNA sequences and suppresses unnecessary transcription. It is also involved in the DNA damage response pathway playing a role in preserving genomic integrity by interacting with proteins such as BRCA1 and 53BP1 involved in DNA repair.

Associated diseases and disorders

HP1 gamma's dysregulation has been associated with cancer and neurological disorders. Overexpression or mutations in this protein can lead to aberrant gene silencing contributing to oncogenesis where cancer-related genes become improperly regulated. Additionally alterations in HP1 gamma function are linked to neurodegenerative diseases potentially through its interaction with proteins like MECP2 whereby changes in the chromatin environment influence neuronal gene expression and function. Therefore understanding HP1 gamma's role provides insights into potential therapeutic targets for these conditions.

Product promise

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11 product images

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
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 - Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 observed at 23 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-HP1 gamma/CBX3 antibody [EPR19803] 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. Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 and Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167)
Predicted band size: 20 kDa
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
All lanes: Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167) at 1/1000 dilution
Lane 1: Human fetal brain lysate at 10 µg
Lane 2: Human fetal kidney lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 20 kDa
Observed band size: 21 kDa
Exposure time: 3min
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1: 3 minutes; Lane 2: 15 seconds.
All lanes: Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167) at 1/1000 dilution
Lane 1: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 2: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 20 kDa
Observed band size: 21 kDa
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: Lanes 1 and 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 30 seconds; Lane 5: 10 seconds; Lanes 6-9: 30 seconds.
All lanes: Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167) at 1/1000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse kidney lysate at 10 µg
Lane 3: Mouse spleen lysate at 10 µg
Lane 4: Rat brain lysate at 10 µg
Lane 5: Rat spleen lysate at 10 µg
Lane 6: C6 (Rat glial tumor cell line) whole cell lysate at 10 µg
Lane 7: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 8: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 9: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 20 kDa
Observed band size: 21 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HP1 gamma/CBX3 with Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling HP1 gamma/CBX3 with Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling HP1 gamma/CBX3 with Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.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 Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (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: Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution. -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.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 Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (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: Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution. -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HP1 gamma/CBX3 with Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal-Isotype control (Rabbit IgG, monoclonal [EPR25A] - 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.
Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
This data was developed using Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HP1 gamma/CBX3 with Anti-HP1 gamma/CBX3 antibody [EPR19803] ab213167 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal - Isotype control (Rabbit IgG, monoclonal [EPR25A] - 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.