重组Anti-Proteasome 20S LMP7抗体[EPR14482(B)] (ab180606)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14482(B)] to Proteasome 20S LMP7
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Proteasome 20S LMP7抗体[EPR14482(B)]
参阅全部 Proteasome 20S LMP7 一抗 -
描述
兔单克隆抗体[EPR14482(B)] to Proteasome 20S LMP7 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details
不适用于: IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: A549, Raji, Jurkat, HeLa and U937 lysates. IHC-P: Human bladder transitional cell carcinoma tissue. ICC/IF: Jurkat and HeLa cells. Flow Cyt (intra): Jurkat cells. IP: Proteasome 20S LMP7 IP in Raji cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR14482(B) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab180606于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IHC-P |
1/1700.
For unpurified use at 1/250 - 1/500. Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) |
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WB |
1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 30 kDa).
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ICC/IF |
1/100.
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Flow Cyt (Intra) |
1/90 - 1/150.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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IHC-P
1/1700. For unpurified use at 1/250 - 1/500. Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) |
WB
1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 30 kDa). |
ICC/IF
1/100. |
Flow Cyt (Intra)
1/90 - 1/150. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
The proteasome is a multicatalytic proteinase complex which is characterized by its ability to cleave peptides with Arg, Phe, Tyr, Leu, and Glu adjacent to the leaving group at neutral or slightly basic pH. The proteasome has an ATP-dependent proteolytic activity. This subunit is involved in antigen processing to generate class I binding peptides. Replacement of PSMB5 by PSMB8 increases the capacity of the immunoproteasome to cleave model peptides after hydrophobic and basic residues. Acts as a major component of interferon gamma-induced sensitivity. Plays a key role in apoptosis via the degradation of the apoptotic inhibitor MCL1. May be involved in the inflammatory response pathway. In cancer cells, substitution of isoform 1 (E2) by isoform 2 (E1) results in immunoproteasome deficiency. -
疾病相关
Defects in PSMB8 are the cause of JMP syndrome (JMPS) [MIM:613732]; also called joint contractures muscular atrophy microcytic anemia and panniculitis-induced lipodystrophy. JBTS1 is an autoinflammatory disorder characterized by childhood onset of joint stiffness and severe contractures of the hands and feet, erythematous skin lesions with subsequent development of severe lipodystrophy, and laboratory evidence of immune dysregulation. Accompanying features include muscle weakness and atrophy, hepatosplenomegaly, and microcytic anemia. -
序列相似性
Belongs to the peptidase T1B family. -
发展阶段
Highly expressed in immature dendritic cells (at protein level). -
翻译后修饰
Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity. -
细胞定位
Cytoplasm. Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 5696 Human
- Omim: 177046 Human
- SwissProt: P28062 Human
- Unigene: 180062 Human
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别名
- ALDD antibody
- D6S216 antibody
- D6S216E antibody
see all
图片
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All lanes : Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/1000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : PSMB8 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 30 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab180606 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.
ab180606 Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267149 (knockout cell lysate ab257130) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab180606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue sections labeling Proteasome 20S LMP7 with purified ab180606 at 1/1700 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Proteasome 20S LMP7 with purified ab180606 at 1:100 dilution (8.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling Proteasome 20S LMP7 with purified ab180606 at 1/90 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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All lanes : Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : PSMB8 knockout A549 cell lysate
Lane 3 : Raji cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 30 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab180606 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.
ab180606 Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267148 (knockout cell lysate ab257129) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab180606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/10000 dilution (Purified) + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 30 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of paraffin-embedded human bladder transitional cell carcinoma tissue labeling Proteasome 20S LMP7 with ab180606 (unpurified) at 1/500 dilution, followed by prediluted ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG. Counterstained with hematoxylin.
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ab180606 (unpurified) staining Proteasome 20S LMP7 in Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.
Negative control: PBS only.
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Immunofluorecenct analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Proteasome 20S LMP7 with ab180606 (unpurified) at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution (left panel). DAPI staining (right panel).
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixedJurkat (Human T cell leukemia cell line from peripheral blood) cells labeling Proteasome 20S LMP7 with ab180606 (unpurified) at 1/150 dilution (red) compared to a Rabbit monoclonal IgG Isotype control (green), followed byGoat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (9)
ab180606 被引用在 9 文献中.
- Zhang L et al. Immunoproteasome subunit β5i promotes perifascicular muscle atrophy in dermatomyositis by upregulating RIG-I. RMD Open 9:N/A (2023). PubMed: 36854567
- Teixeira VON et al. The role of proteasome in muscle wasting of experimental arthritis. Adv Rheumatol 63:14 (2023). PubMed: 36949513
- Sun HT et al. Quercetin suppresses inflammatory cytokine production in rheumatoid arthritis fibroblast-like synoviocytes. Exp Ther Med 22:1260 (2021). PubMed: 34603528
- Tian W et al. A Novel Prognostic Tool for Glioma Based on Enhancer RNA-Regulated Immune Genes. Front Cell Dev Biol 9:798445 (2021). PubMed: 35127714
- Du SH et al. Co-Inhibition of the Immunoproteasome Subunits LMP2 and LMP7 Ameliorates Immune Thrombocytopenia. Front Immunol 11:603278 (2020). PubMed: 33552061
- Woodle ES et al. Proteasomal adaptations underlying carfilzomib-resistance in human bone marrow plasma cells. Am J Transplant 20:399-410 (2020). PubMed: 31595669
- Liu Y et al. miR-451a is downregulated and targets PSMB8 in prostate cancer. Kaohsiung J Med Sci 36:494-500 (2020). PubMed: 32128987
- Javitt A et al. Pro-inflammatory Cytokines Alter the Immunopeptidome Landscape by Modulation of HLA-B Expression. Front Immunol 10:141 (2019). PubMed: 30833945
- Xie X et al. The immunoproteasome catalytic ß5i subunit regulates cardiac hypertrophy by targeting the autophagy protein ATG5 for degradation. Sci Adv 5:eaau0495 (2019). PubMed: 31086810