Name: Anti-HP1 gamma/CBX3 antibody [EPR19803]
Brand: RabMAb
SKU: ab213167
Rating: 4 (3 reviews)

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Western blot - 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

Carrier Free

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

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19803
Isotype

IgG
Research areas

Alternative Versions
- Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant Human HP1 gamma/CBX3 protein (ab124548)

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
WB	(1)	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)	1/100.

Notes
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.

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
modifications

Phosphorylated 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.
Target information above from: UniProt accession Q13185 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 458234 Mouse
- Unigene: 485141 Mouse
see all
Alternative names
- CBX 3 antibody
- CBX3 antibody
- CBX3_HUMAN antibody
- Chromobox homolog 3 (HP1 gamma homolog, Drosophila) antibody
- Chromobox homolog 3 antibody
- Chromobox protein homolog 3 antibody
- GAMMA antibody
- HECH antibody
- Heterochromatin like protein 1 antibody
- Heterochromatin protein 1 homolog gamma antibody
- Heterochromatin protein HP1 gamma antibody
- HP1 gamma antibody
- HP1 gamma homolog antibody
- HP1Hs gamma antibody
- Modifier 2 protein antibody
see all

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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.
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 3 minutes; Lane 2: 15 seconds.
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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 kDa

Blocking/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.
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.
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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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.
Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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.
Flow Cytometry (Intracellular) - Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

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.
Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab213167? Please let us know so that we can cite the reference in this datasheet.

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

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1-3 of 3 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (293T)

Gel Running Conditions

Reduced Denaturing (4-20% Gradiant)

Loading amount

20 µg

Specification

293T

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 25 2017

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Epithelial Cells)

Permeabilization

Yes - 0.2% TritonX-100

Specification

Epithelial Cells

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 28 2016

Application

ChIP

Sample

Human Cell lysate - nuclear (B Cells)

Negative control

Isotype IgG

Specification

B Cells

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde

Positive control

Specific Gene promoter

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 01 2016

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (3)

Customer reviews and Q&As

Western blot abreview for Anti-HP1 gamma/CBX3 antibody [EPR19803]

Immunocytochemistry/ Immunofluorescence abreview for Anti-HP1 gamma/CBX3 antibody [EPR19803]

ChIP abreview for Anti-HP1 gamma/CBX3 antibody [EPR19803]

Post-translational
modifications