Overview

  • Product name

    Anti-HP1 alpha antibody - ChIP Grade
    See all HP1 alpha primary antibodies
  • Description

    Goat polyclonal to HP1 alpha - ChIP Grade
  • Host species

    Goat
  • Tested applications

    Suitable for: ICC/IF, IP, IHC-P, ChIP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog, Rhesus monkey, Chinese hamster
  • Immunogen

    Synthetic peptide:

    C-NKRKSNFSNSADDIK

    , corresponding to internal sequence amino acids 88-102 of Human HP1 alpha (NP_036249.1).

  • Positive control

    • Recombinant Human HP1 alpha protein (ab124602) can be used as a positive control in WB. Human fibroblast cell lines A431 and HeLa cell lysates. Human uterus tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab77256 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
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 2 - 3 µg/ml.
ChIP Use at an assay dependent dilution. PubMed: 20663865
WB Use a concentration of 0.03 - 0.1 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

Target

  • Function

    Component of heterochromatin that recognizes and binds histone H3 tails methylated at 'Lys-9' (H3K9me), leading to epigenetic repression. In contrast, it is excluded from chromatin when 'Tyr-41' of histone H3 is phosphorylated (H3Y41ph). Can interact with lamin-B receptor (LBR). This interaction can contribute to the association of the heterochromatin with the inner nuclear membrane. Involved in the formation of functional kinetochore through interaction with MIS12 complex proteins.
  • Sequence similarities

    Contains 2 chromo domains.
  • Post-translational
    modifications

    Phosphorylation of HP1 and LBR may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle (By similarity). Phosphorylated during interphase and possibly hyper-phosphorylated during mitosis.
    Ubiquitinated.
  • Cellular localization

    Nucleus. Chromosome. Chromosome > centromere. Component of centromeric and pericentromeric heterochromatin. Associates with chromosomes during mitosis. Associates specifically with chromatin during metaphase and anaphase.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen p25 antibody
    • CBX5 antibody
    • CBX5_HUMAN antibody
    • CG8409 antibody
    • Chromobox 5 antibody
    • Chromobox homolog 5 (HP1 alpha homolog, Drosophila) antibody
    • Chromobox homolog 5 antibody
    • Chromobox protein homolog 5 antibody
    • Epididymis luminal protein 25 antibody
    • HEL25 antibody
    • Heterochromatin protein 1 alpha antibody
    • Heterochromatin protein 1 antibody
    • Heterochromatin protein 1 homolog alpha antibody
    • HP1 alpha antibody
    • HP1 alpha homolog antibody
    • HP1 antibody
    • HP1A antibody
    • HP1Hs alpha antibody
    • Su(var)205 antibody
    see all

Images

  • All lanes : Anti-HP1 alpha antibody - ChIP Grade (ab77256) at 0.3 µg/ml

    Lane 1 : A431 cell line lysates in RIPA buffer
    Lane 2 : HeLa cell line lysates in RIPA buffer
    Lane 3 : NIH3T3 cell line lysates in RIPA buffer

    Lysates/proteins at 35 µg per lane.

    Predicted band size: 22 kDa



    Primary incubation was 1 hour. Detected by chemiluminescence.

  • ab77256 at 2.5µg/ml staining HP1 alpha in human uterus tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded sections). Tissue underwent antigen retrieval in steam with citrate buffer (pH 6.0). The AP-staining procedure was used for detection.

  • ab77256 staining HP1 alpha in Mouse 3T3 fibroblasts by Immunocytochemistry/ Immunoflourescence. Cells were PFA-fixed and permeabilized in 0.2% Triton X-100 in PBS. The primary antibody was diluted 1/100 in PBS + Tween 1:1000 and incubated with the sample for 30 minutes. The secondary antibody was Cy2®-conjugated Donkey anti-Goat polyclonal, diluted 1/100.

    See Abreview

  • Anti-HP1 alpha antibody - ChIP Grade (ab77256) at 0.03 µg/ml + HeLa lysate in RIPA buffer at 35 µg

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    Primary incubation was 1 hour. Detected by chemiluminescence.
  • ab77256 (2µg/ml) staining of paraffin embedded Human Uterus. Steamed antigen retrieval with citrate buffer pH 6, HRP-staining.

References

This product has been referenced in:

  • Hervás-Corpión I  et al. Early alteration of epigenetic-related transcription in Huntington's disease mouse models. Sci Rep 8:9925 (2018). Read more (PubMed: 29967375) »
  • Chow TT  et al. Local enrichment of HP1alpha at telomeres alters their structure and regulation of telomere protection. Nat Commun 9:3583 (2018). Read more (PubMed: 30181605) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 12 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

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

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

Abcam user community

Verified customer

Submitted Nov 28 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde

Ahmar Aziz

Verified customer

Submitted May 03 2016

Application
Western blot
Sample
Rat Cell lysate - whole cell (Epithelial cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
75 µg
Specification
Epithelial cells
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Nov 11 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HeLa, RPE-1)
Specification
HeLa, RPE-1
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 08 2013

Question
Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

Questions - ChIP

Sample type, species
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)
Controls used (positive/negative antibodies, positive/negative control regions, no template control in PCR)
Lot number/order number/date of purchase



WB

Sample preparation (cell type, species, lysis buffer, protease inhibitors, phosphatase inhibitors)
Amount of protein loaded
Electrophoresis and transfer conditions
Blocking conditions
Primary antibody conditions (diluent/dilution, incubation time/temperature)
Secondary antibody conditions (diluent/dilution, incubation time/temperature)
Wash steps and buffer
Detection method
Controls used (sample, alternative primary ab, no primary)



Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More
Application
Immunoprecipitation
Sample
Human Cell lysate - nuclear (HEK-293)
Total protein in input
1000 µg
Specification
HEK-293
Immuno-precipitation step
Protein G

Abcam user community

Verified customer

Submitted Mar 07 2011

Application
Western blot
Sample
Human Cell lysate - nuclear (HEK-293)
Loading amount
200000 cells
Specification
HEK-293
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Benny Garfinkel

Verified customer

Submitted Feb 03 2011

1-10 of 12 Abreviews or Q&A

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