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Product Info Summary
| SKU: | M02780-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-HP1 alpha/CBX5 Antibody Picoband® (monoclonal, 8G6)
SKU/Catalog Number
M02780-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-HP1 alpha/CBX5 Antibody Picoband® (monoclonal, 8G6) catalog # M02780-2. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-HP1 alpha/CBX5 Antibody Picoband® (monoclonal, 8G6) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M02780-2)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Monoclonal
Clone Number
8G6
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human HP1 alpha/CBX5 recombinant protein (Position: M1-S191).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M02780-2 is reactive to CBX5 in Human, Mouse, Rat
Observed Molecular Weight
22 kDa
Calculated molecular weight
22.2 kDa
Background of CBX5
This gene encodes a highly conserved nonhistone protein, which is a member of the heterochromatin protein family. The protein is enriched in the heterochromatin and associated with centromeres. The protein has a single N-terminal chromodomain which can bind to histone proteins via methylated lysine residues, and a C-terminal chromo shadow-domain (CSD) which is responsible for the homodimerization and interaction with a number of chromatin-associated nonhistone proteins. The encoded product is involved in the formation of functional kinetochore through interaction with essential kinetochore proteins. The gene has a pseudogene located on chromosome 3. Multiple alternatively spliced variants, encoding the same protein, have been identified.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M02780-2 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human U87 whole cell, human K562 whole cell, human Jurkat whole cell, rat brain tissue, rat PC-12 whole cell, mouse brain tissue, mouse SP2/0 whole cell
IHC: rat brain tissue, human adrenocortical adenoma tissue, human ovarian serous adenocarcinoma tissue, human breast cancer tissue, human gallbladder adenocarcinoma tissue, human lymphoma tissue, human rectal cancer tissue, human renal clear cell carcinoma tissue, mouse brain tissue
ICC/IF: A549 cell
FCM: U251 cell
Validation Images & Assay Conditions
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Western blot analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
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 30ug of sample under reducing conditions.
Lane 1: human U87 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: 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 mouse anti-HP1 alpha/CBX5 antigen affinity purified monoclonal antibody (Catalog # M02780-2) 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-mouse 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 # EK1001) 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.
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IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of rat brain 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human adrenocortical adenoma 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human breast 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human gallbladder adenocarcinoma 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human lymphoma 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human rectal 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of human renal clear cell carcinoma 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IHC analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in paraffin-embedded section of mouse brain 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 2μg/ml mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
IF analysis of HP1 alpha/CBX5 using anti-HP1 alpha/CBX5 antibody (M02780-2).
HP1 alpha/CBX5 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti-HP1 alpha/CBX5 Antibody (M02780-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.
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Flow Cytometry analysis of U251 cells using anti-HP1 alpha/CBX5 antibody (M02780-2).
Overlay histogram showing U251 cells stained with M02780-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- HP1 alpha/CBX5 Antibody (M02780-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-HP1 alpha/CBX5 Antibody Picoband® (monoclonal, 8G6) (M02780-2)
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