Anti-PSME1 Antibody Picoband™

Product Name

Anti-PSME1 Antibody Picoband™

View all PA28 Activator alpha Subunit/PSME1 Antibodies

SKU/Catalog Number

A04638-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-PSME1 Antibody Picoband™ catalog # A04638-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

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 freezing and thawing.

Cite This Product

Anti-PSME1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04638-2)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human PSME1 recombinant protein (Position: M1-Y249).

*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

A04638-2 is reactive to PSME1 in Human, Mouse, Rat

Applications

A04638-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

30 kDa

Calculated molecular weight

28.723kDa

Background of PA28 Activator alpha Subunit/PSME1

Proteasome activator complex subunit 1 is a protein that in humans is encoded by the PSME1 gene. The 26S proteasome is a multicatalytic proteinase complex with a highly ordered structure composed of 2 complexes, a 20S core and a 19S regulator. The 20S core is composed of 4 rings of 28 non-identical subunits; 2 rings are composed of 7 alpha subunits and 2 rings are composed of 7 beta subunits. The 19S regulator is composed of a base, which contains 6 ATPase subunits and 2 non-ATPase subunits, and a lid, which contains up to 10 non-ATPase subunits. Proteasomes are distributed throughout eukaryotic cells at a high concentration and cleave peptides in an ATP/ubiquitin-dependent process in a non-lysosomal pathway. An essential function of a modified proteasome, the immunoproteasome, is the processing of class I MHC peptides. The immunoproteasome contains an alternate regulator, referred to as the 11S regulator or PA28, that replaces the 19S regulator. Three subunits (alpha, beta and gamma) of the 11S regulator have been identified. This gene encodes the alpha subunit of the 11S regulator, one of the two 11S subunits that is induced by gamma-interferon. Three alpha and three beta subunits combine to form a heterohexameric ring. Alternative splicing results in multiple transcript variants.

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.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

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, 0.1-0.25 µg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of PSME1 using anti-PSME1 antibody (A04638-2).
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 30 ug of sample under reducing conditions.
Lane 1: human Raji whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat spleen tissue lysates,
Lane 5: rat RH35 whole cell lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-PSME1 antigen affinity purified polyclonal antibody (Catalog # A04638-2) at 0.25 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PSME1 at approximately 30 kDa. The expected band size for PSME1 is at 29 kDa.

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Figure 2. IF analysis of PSME1 using anti-PSME1 antibody (A04638-2).
PSME1 was detected in an 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 5 μg/mL rabbit anti-PSME1 Antibody (A04638-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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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Figure 3. Flow Cytometry analysis of JK cells using anti-PSME1 antibody (A04638-2).
Overlay histogram showing JK cells stained with A04638-2 (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 rabbit anti-PSME1 Antibody (A04638-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For PSME1 (Source: Uniprot.org, NCBI)

Alternative Names

11S regulator complex alpha subunit; 11S Regulator Complex Subunit alpha; 29-kD MCP activator subunit; IFI5111; IFI5111MGC8628; IGUP I-5111; Interferon gamma up-regulated I-5111 protein; interferon-gamma IEF SSP 5111; interferon-gamma-inducible protein 5111; PA28 Activator alpha Subunit; PA28A; PA28alphaActivator of multicatalytic protease subunit 1; proteasome (prosome, macropain) activator subunit 1 (PA28 alpha); Proteasome activator 28 subunit alpha; proteasome activator complex subunit 1; proteasome activator subunit-1; PSME1; REG alpha; REGalpha; REG-alpha PSME1 HEL-S-129m, IFI5111, PA28A, PA28alpha, REGalpha proteasome activator subunit 1 proteasome activator complex subunit 1|11S regulator complex subunit alpha|29-kD MCP activator subunit|IGUP I-5111|activator of multicatalytic protease subunit 1|epididymis secretory sperm binding protein Li 129m|interferon gamma up-regulated I-5111 protein|interferon-gamma IEF SSP 5111|interferon-gamma-inducible protein 5111|proteasome (prosome, macropain) activator subunit 1 (PA28 alpha)

For more info on PSME1, check out the PSME1 Infographic

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In this infographic, you will see the following information for PSME1: 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].

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