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Catalog Number | orb654408 |
---|---|
Category | Antibodies |
Description | Anti-HP1 alpha/CBX5 Antibody. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human HP1 alpha/CBX5 recombinant protein (Position: M1-S191). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 22 kDa |
UniProt ID | P45973 |
Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
Alternative names | CD3 antigen, epsilon polypeptide; CD3e molecule; C Read more... |
Note | For research use only |
Application notes | Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 4μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human U-87MG whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human Raji whole cell lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: mouse SP2/0 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HP1 alpha/CBX5 antigen affinity purified polyclonal antibody (Catalog # orb654408) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HP1 alpha/CBX5 at approximately 22KD. The expected band size for HP1 alpha/CBX5 is at 22KD.
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IF analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (orb654408). HP1 alpha/CBX5 was detected in immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-HP1 alpha/CBX5 Antibody (orb654408) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of K562 cells using anti-HP1 alpha/CBX5 antibody (orb654408). Overlay histogram showing K562 cells stained with orb654408 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HP1 alpha/CBX5 Antibody (orb654408, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, ICC, IF, IHC, IP, WB | |
Human, Mouse, Rat | |
Rabbit | |
Monoclonal | |
Unconjugated |