HP1 alpha Recombinant Rabbit Monoclonal Antibody [SR44-03]
cat.: ET1602-8
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SR44-03
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 22 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human HP1 alpha aa 101-191 / 191.
Positive control: HeLa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, Hela, MCF-7, human tonsil tissue, human breast tissue, rat uterus tissue, human uterus tissue, mouse uterus tissue, SH-SY5Y.
Subcellular location: Nucleus, Chromosome
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P45973 Human | Q61686 Mouse
Entrez Gene: 300266 Rat
Alternative names: Antigen p25 CBX5 CBX5_HUMAN CG8409 Chromobox 5 Chromobox homolog 5 (HP1 alpha homolog, Drosophila) Chromobox homolog 5 Chromobox protein homolog 5 Epididymis luminal protein 25 HEL25 Heterochromatin protein 1 alpha Heterochromatin protein 1 Heterochromatin protein 1 homolog alpha HP1 alpha HP1 alpha homolog HP1 HP1A HP1Hs alpha Su(var)205
Images
ET1602-8_1.jpg Fig1: Western blot analysis of HP1 alpha on different lysates with Rabbit anti-HP1 alpha antibody (ET1602-8) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: MCF7 cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: RAW264.7 cell lysate (10 µg/Lane)
Lane 5: C6 cell lysate (10 µg/Lane)
Lane 6: PC-12 cell lysate (10 µg/Lane)
Lane 7: Mouse brain tissue lysate (20 µg/Lane)

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

Exposure time: 24 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1602-8) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1602-8_2.jpg Fig2: ICC staining of HP1 alpha in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-8, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1602-8_3.jpg Fig3: ICC staining of HP1 alpha in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-8, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1602-8_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-HP1 alpha antibody (ET1602-8) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-8) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1602-8_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-HP1 alpha antibody (ET1602-8) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-8) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1602-8_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat uterus tissue with Rabbit anti-HP1 alpha antibody (ET1602-8) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-8) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1602-8_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-HP1 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-8, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1602-8_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse uterus tissue using anti-HP1 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-8, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1602-8_9.jpg Fig9: Flow cytometric analysis of HP1 alpha was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1602-8, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.