Key features and details
- Rabbit polyclonal to Proteasome 20S LMP7
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
存储溶液Constituents: 0.1% BSA, 99% PBS
Concentration information loading...
纯度Immunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab3329 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 20 kDa.Can be blocked with Human Proteasome 20S LMP7 peptide (ab4945).|
|IHC-P||Use a concentration of 1 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
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.
- Information by UniProt
- ALDD antibody
- D6S216 antibody
- D6S216E antibody
All lanes : Anti-Proteasome 20S LMP7 antibody (ab3329) at 2 µg/ml
Lane 1 : HeLa whole cell extracts
Lane 2 : HeLa treated with IFN gamma (100ng/ml IFN gamma for 72h) whole cell extracts
Lane 3 : U-937 whole cell extracts
Lane 4 : Raji whole cell extracts
Lane 5 : Ramos whole cell extracts
Lysates/proteins at 30 µg per lane.
All lanes : Goat anti-Rabbit IgG (H+L) HRP conjugate at 1/2500 dilution
Observed band size: 20 kDa why is the actual band size different from the predicted?
Detected by chemiluminescence.
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Proteasome 20S LMP7 with ab3329 at 5µg/ml. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with or without Anti-Proteasome 20S LMP7 antibody (ab3329), at a concentration of 5µg/ml for 1 hour at room temperature, and then incubated with a Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody at a dilution of 1/1000 for 1 hour s at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.
Ab3329 staining Human normal skin. Staining is localized to cytoplasmic and nuclear compartments.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required
Western blot of proteasome 20S LMP7 from HeLa cell extract using ab3329.
ICC/IF image of ab3329 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3329, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab3329 被引用在 33 文献中.
- Chen C et al. Resveratrol as a new inhibitor of immunoproteasome prevents PTEN degradation and attenuates cardiac hypertrophy after pressure overload. Redox Biol 20:390-401 (2019). PubMed: 30412827
- Li FD et al. Ablation and Inhibition of the Immunoproteasome Catalytic Subunit LMP7 Attenuate Experimental Abdominal Aortic Aneurysm Formation in Mice. J Immunol 202:1176-1185 (2019). PubMed: 30642978
- Lee MJ et al. H727 cells are inherently resistant to the proteasome inhibitor carfilzomib, yet require proteasome activity for cell survival and growth. Sci Rep 9:4089 (2019). PubMed: 30858500
- 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
- Jimenez-Guardeño JM et al. Immunoproteasome activation enables human TRIM5a restriction of HIV-1. Nat Microbiol 4:933-940 (2019). PubMed: 30886358