Product Info Summary
SKU: | M00409-2 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, IHC-F, ICC, WB |
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Product info
Product Name
Anti-KAP1/TRIM28 Antibody Picoband® (monoclonal, 9E3)
SKU/Catalog Number
M00409-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-KAP1/TRIM28 Antibody Picoband® (monoclonal, 9E3) catalog # M00409-2. Tested in Flow Cytometry, IF, IHC, IHC-F, 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-KAP1/TRIM28 Antibody Picoband® (monoclonal, 9E3) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00409-2)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
9E3
Isotype
Mouse IgG2a
Immunogen
E.coli-derived human KAP1 recombinant protein (Position: A699-P835). Human KAP1 shares 94.9% amino acid (aa) sequence identity with both mouse and rat KAP1.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00409-2 is reactive to TRIM28 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
100 kDa
Calculated molecular weight
88.55kDa
Background of KAP1
Tripartite motif-containing 28 (TRIM28), also known as transcriptional intermediary factor 1β (TIF1β) and KAP1 (KRAB-associated protein-1), is a protein that in humans is encoded by the TRIM28 gene. The protein encoded by this gene mediates transcriptional control by interaction with the Kruppel-associated box repression domain found in many transcription factors. The protein localizes to the nucleus and is thought to associate with specific chromatin regions. KAP1 is a ubiquitously expressed protein involved in many critical functions including: transcriptional regulation, cellular differentiation and proliferation, DNA damage repair, viral suppression, and apoptosis. Its functionality is dependent upon post-translational modifications. Phosphorylation of KAP1 acts as a deactivator of the protein in many of its mechanisms while sumoylation acts as an activator.
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
M00409-2 is guaranteed for Flow Cytometry, IF, IHC, IHC-F, 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell,, human PC-3 whole cell,, human HEK293 whole cell,, human A549 whole cell,, human Jurkat whole cell,, human THP-1 whole cell,, rat PC-12 whole cell,, mouse NIH/3T3 whole cell
IHC: human mammary cancer tissue, human rectal cancer tissue, rat intestine tissue
IHC-F: human placenta tissue
ICC/IF: U20S cell
IF: human rectal cancer tissue
FCM: A549 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
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 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates;
Lane 2: human PC-3 whole cell lysates;
Lane 3: human HEK293 whole cell lysates;
Lane 4: human A549 whole cell lysates;
Lane 5: human Jurkat whole cell lysates;
Lane 6: human THP-1 whole cell lysates;
Lane 7: rat PC-12 whole cell lysates;
Lane 8: mouse NIH/3T3 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-KAP1/TRIM28 antigen affinity purified monoclonal antibody (Catalog # M00409-2) 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 KAP1/TRIM28 at approximately 100KD. The expected band size for KAP1/TRIM28 is at 100KD.
Click image to see more details
Figure 2. IHC analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 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 mouse anti-KAP1/TRIM28 Antibody (M00409-2) 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
Figure 3. IHC analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 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 1μg/ml mouse anti-KAP1/TRIM28 Antibody (M00409-2) 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
Figure 4. IHC analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 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 mouse anti-KAP1/TRIM28 Antibody (M00409-2) 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
Figure 5. IF analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 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-KAP1/TRIM28 Antibody (M00409-2) 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. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 6. IF analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 was detected in immunocytochemical section of U20S 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 2μg/mL mouse anti-KAP1/TRIM28 Antibody (M00409-2) 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.
Click image to see more details
Figure 7. Flow Cytometry analysis of A549 cells using anti- KAP1/TRIM28 antibody (M00409-2).
Overlay histogram showing A549 cells stained with M00409-2 (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- KAP1/TRIM28 Antibody (M00409-2, 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.
Click image to see more details
Figure 8. IHC analysis of KAP1/TRIM28 using anti-KAP1/TRIM28 antibody (M00409-2).
KAP1/TRIM28 was detected in frozen section of human placenta tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-KAP1/TRIM28 Antibody (M00409-2) 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.
Protein Target Info & Infographic
Gene/Protein Information For TRIM28 (Source: Uniprot.org, NCBI)
Gene Name
TRIM28
Full Name
Transcription intermediary factor 1-beta
Weight
88.55kDa
Superfamily
TRIM/RBCC family
Alternative Names
E3 SUMO protein ligase TRIM28 antibody; E3 SUMO-protein ligase TRIM28 antibody; FLJ29029 antibody; KAP 1 antibody; KAP-1 antibody; KRAB associated protein 1 antibody; KRAB interacting protein 1 antibody; KRAB-associated protein 1 antibody; KRAB-interacting protein 1 antibody; KRIP 1 antibody; KRIP-1 antibody; KRIP1 antibody; Nuclear corepressor KAP 1 antibody; Nuclear corepressor KAP-1 antibody; RING finger protein 96 antibody; RNF96 antibody; TF1B antibody; TIF1 beta antibody; TIF1-beta antibody; TIF1B antibody; TIF1B_HUMAN antibody; Transcription intermediary factor 1 beta antibody; Transcription intermediary factor 1-beta antibody; Trim28 antibody; Tripartite motif containing 28 antibody; tripartite motif containing protein 28 antibody; Tripartite motif-containing protein 28 antibody TRIM28 KAP1, PPP1R157, RNF96, TF1B, TIF1B tripartite motif containing 28 transcription intermediary factor 1-beta|E3 SUMO-protein ligase TRIM28|KAP-1|KRAB [Kruppel-associated box domain]-associated protein 1|KRAB-interacting protein 1|KRIP-1|RING finger protein 96|RING-type E3 ubiquitin transferase TIF1-beta|TIF1-beta|nuclear corepressor KAP-1|protein phosphatase 1, regulatory subunit 157|transcriptional intermediary factor 1-beta
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on TRIM28, check out the TRIM28 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for TRIM28: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-KAP1/TRIM28 Antibody Picoband® (monoclonal, 9E3) (M00409-2)
Hello CJ!
M00409-2 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Hao Y,Bai S,Peng J,Hong R,Ding J,Li Z,Guan Y.TRIM27-mediated ubiquitination of PPARγ promotes glutamate-induced cell apoptosis and inflammation.Exp Cell Res.2020 Dec 29:112437.doi:10.1016/j.yexcr.2020. 112437.Epub ahead of print.PMID:33385414.
Species: Human
M00409-2 usage in article: APP:WB, SAMPLE:HT22 CELL, DILUTION:NA
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