Overview

  • Product name
  • Description
    Rabbit polyclonal to HP1 gamma
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse HP1 gamma.

    Read Abcam's proprietary immunogen policy .

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; MCF7; HEK293.

Properties

Applications

Our Abpromise guarantee covers the use of ab10480 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 1/500 - 1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 21 kDa).

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: HP1 gamma knockout HAP1 cell lysate (20 µg)
    Lane 3: NIH/3T3 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab10480 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab10480 was shown to recognize HP1 gamma when HP1 gamma knockout samples were used, along with additional cross-reactive bands. Wild-type and HP1 gamma knockout samples were subjected to SDS-PAGE. ab10480 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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 : Anti-HP1 gamma antibody (ab10480) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 24 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 49 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • Ab10480 recognizes the tagged recombinant HP1 gamma protein (ab88011) which has an expected molecular weight of ~50 kDa.
  • Ab10480 recognizes the tagged recombinant HP1 gamma protein (ab88011) which has an expected molecular weight of ~50 kDa.

References

This product has been referenced in:
  • Sherrard A  et al. Streamlined histone-based fluorescence lifetime imaging microscopy (FLIM) for studying chromatin organisation. Biol Open 7:N/A (2018). Read more (PubMed: 29535103) »
  • Magaraki A  et al. Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development. Epigenetics Chromatin 10:11 (2017). IF ; Mouse . Read more (PubMed: 28293300) »
See all 13 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - nuclear (293T)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
15 µg
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 16 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mouse 14.5dpc)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris pH 9
Permeabilization
No
Specification
Mouse 14.5dpc
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 15 2013

Answer

Thank you for contacting us.

I am attaching few questions here, please answers these as these will help us to investigate what went wrong with these antibodies;

Sample preparation (X-ChIP/N-ChIP, cross-linking conditions, DNA fragment size)
Immunoprecipitation conditions (buffer, amount of detergent, primary antibody manufacturer/dilution, incubation time, matrix for isolating antibody complex)
Wash steps
DNA purification (decrosslinking conditions, DNA purification method)
DNA detection method (regions amplified in PCR, microarray, direct sequencing
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question
Answer

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I am very pleased to hear you would like to accept our offer and test ab10480in mouse tissue. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse tissueand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin, epidermis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
Yes - TBST
Specification
Skin, epidermis
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 16 2010

Application
Immunocytochemistry
Sample
Human Cultured Cells (Colorectal Cancer Cells)
Permeabilization
Yes - Triton X-100
Specification
Colorectal Cancer Cells
Blocking step
Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 06 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Colorectal Cancer Cells)
Gel Running Conditions
Reduced Denaturing (12% Gel)
Loading amount
200 µg
Specification
Colorectal Cancer Cells
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Jan 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human Colon Cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: DAKO Target Retrieval Solution
Permeabilization
No
Specification
Human Colon Cancer
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HMEC human mammary epithelial cells)
Permeabilization
Yes - fixation was combined with permeabilazation with 4%PF in PBS 0,1% Triton X-100
Specification
HMEC human mammary epithelial cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Silvia Sirchia

Verified customer

Submitted Jun 09 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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