AB251540

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

RabMAb
Recombinant
KO Validated
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(1 Publication)

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. Cited in 1 publication.

View Alternative Names

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

11 Images

Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using 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 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

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.

Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using 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 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.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

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.

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using 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 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.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using 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 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.

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.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

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.

Supplier Data

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

This data was developed using 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] (<a href='/en-us/products/primary-antibodies/hp1-gamma-cbx3-antibody-epr19803-ab213167'>ab213167</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 20 kDa

Observed band size: 21 kDa

false

Supplier Data

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

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (<a href='/en-us/products/primary-antibodies/hp1-gamma-cbx3-antibody-epr19803-ab213167'>ab213167</a>) 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

false

Exposure time: 3min

Lab

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

This data was developed using 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 - 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.

All lanes:

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (<a href='/en-us/products/primary-antibodies/hp1-gamma-cbx3-antibody-epr19803-ab213167'>ab213167</a>)

Predicted band size: 20 kDa

false

Supplier Data

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

This data was developed using 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] (<a href='/en-us/products/primary-antibodies/hp1-gamma-cbx3-antibody-epr19803-ab213167'>ab213167</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 20 kDa

Observed band size: 21 kDa

false

Unconjugated

Anti-HP1 gamma/CBX3 antibody [EPR19803]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR19803

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

ICC/IF, IHC-P, WB, Flow Cyt (Intra)

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "This application is recommended for human and mouse only." }, "Mouse": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "This application is recommended for human and mouse only." }, "Rat": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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Product details

ab251540 is the carrier-free version of ab213167.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

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

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.

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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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).

See full target information CBX3

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Cancer cell 39:1479-1496.e18 PubMed34653364

2021

Signatures of plasticity, metastasis, and immunosuppression in an atlas of human small cell lung cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Joseph M Chan,Álvaro Quintanal-Villalonga,Vianne Ran Gao,Yubin Xie,Viola Allaj,Ojasvi Chaudhary,Ignas Masilionis,Jacklynn Egger,Andrew Chow,Thomas Walle,Marissa Mattar,Dig V K Yarlagadda,James L Wang,Fathema Uddin,Michael Offin,Metamia Ciampricotti,Besnik Qeriqi,Amber Bahr,Elisa de Stanchina,Umesh K Bhanot,W Victoria Lai,Matthew J Bott,David R Jones,Arvin Ruiz,Marina K Baine,Yanyun Li,Natasha Rekhtman,John T Poirier,Tal Nawy,Triparna Sen,Linas Mazutis,Travis J Hollmann,Dana Pe'er,Charles M Rudin

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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

Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (AB251540)

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

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

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

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

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Primary Antibodies

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

Product promise