Rabbit TBP-1/PSMC3 Polyclonal Antibody | anti-PSMC3 antibody

Anti-TBP-1/PSMC3 Antibody

Cat. Num. AAA19314

• Gene Names: PSMC3; TBP1
• Reactivity: Human, Mouse, Rat
• Applications: Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
• Purity: Immunogen affinity purified.
• Synonyms: TBP-1/PSMC3; Polyclonal Antibody; Anti-TBP-1/PSMC3 Antibody; PSMC3; TBP1; 26S proteasome regulatory subunit 6A; 26S proteasome AAA-ATPase subunit RPT5; Proteasome 26S subunit ATPase 3; Proteasome subunit P50; Tat-binding protein 1; TBP-1; proteasome 26S subunit; ATPase 3; anti-PSMC3 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Rabbit IgG polyclonal antibody for TBP-1/PSMC3 detection.
• Purity/Purification: Immunogen affinity purified.
• Form/Format: Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

• Applicable Applications for anti-PSMC3 antibody: Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
• Application Notes: WB: 0.1-0.25ug/ml|Human, Mouse, Rat|; IHC-P: 2-5ug/ml|Human, Rat|; FC/FACS/FCM: 1-3ug/1x106 cells|Human|; Direct ELISA: 0.1-0.5ug/ml|Human|
• Immunogen: E Coli-derived human TBP-1/PSMC3 recombinant protein (Position: M1-A439).
• Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Recommended Detection Systems: Recommended Detection Systems
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 7. Flow Cytometry analysis of 293T cells using anti-TBP-1/PSMC3 antibody (AAA19314).Overlay histogram showing 293T cells stained with AAA19314 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TBP-1/PSMC3 Antibody (AAA19314, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistchemistry)

(Figure 6. IHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).TBP-1/PSMC3 was detected in paraffin-embedded section of human pancreatic 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 rabbit anti-TBP-1/PSMC3 Antibody (AAA19314) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).TBP-1/PSMC3 was detected in paraffin-embedded section of human bladder 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 rabbit anti-TBP-1/PSMC3 Antibody (AAA19314) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).TBP-1/PSMC3 was detected in paraffin-embedded section of human renal 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 rabbit anti-TBP-1/PSMC3 Antibody (AAA19314) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).TBP-1/PSMC3 was detected in paraffin-embedded section of human appendicitis 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 rabbit anti-TBP-1/PSMC3 Antibody (AAA19314) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).TBP-1/PSMC3 was detected in paraffin-embedded section of rat cardiac muscle 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 rabbit anti-TBP-1/PSMC3 Antibody (AAA19314) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of TBP-1/PSMC3 using anti-TBP-1/PSMC3 antibody (AAA19314).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 HEK293 whole cell lysatesLane 2: human HELA whole cell lysatesLane 3: human HEPG2 whole cell lysatesLane 4: human MCF-7 whole cell lysatesLane 5: human U20S whole cell lysatesLane 6: human PC-3 whole cell lysatesLane 7: human U87 whole cell lysatesLane 8: rat stomach tissue lysatesLane 9: mouse lung tissue 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 rabbit anti-TBP-1/PSMC3 antigen affinity purified polyclonal antibody (Catalog # AAA19314) at 0. 25 μg/mL overnight at 4 degree 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 # with Tanon 5200 system. A specific band was detected for TBP-1/PSMC3 at approximately 50KD. The expected band size for TBP-1/PSMC3 is at 50KD.)

Related Product Information for anti-PSMC3 antibody

26S protease regulatory subunit 6A, also known as 26S proteasome AAA-ATPase subunit Rpt5, is an enzyme that in humans is encoded by the PSMC3 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. This gene encodes one of the ATPase subunits, a member of the triple-A family of ATPases that have chaperone-like activity. This subunit may compete with PSMC2 for binding to the HIV tat protein to regulate the interaction between the viral protein and the transcription complex. A pseudogene has been identified on chromosome 9.

References

1. Corn, P. G., McDonald, E. R., III, Herman, J. G., El-Deiry, W. S. Tat-binding protein-1, a component of the 26S proteasome, contributes to the E3 ubiquitin ligase function of the von Hippel-Lindau protein. Nature Genet. 35: 229-237, 2003.
2. DeMartino, G. N., Proske, R. J., Moomaw, C. R., Strong, A. A., Song, X., Hisamatsu, H., Tanaka, K., Slaughter, C. A. Identification, purification, and characterization of a PA700-dependent activator of the proteasome. J. Biol. Chem. 271: 3112-3118, 1996.
3. Hoyle, J., Tan, K. H., Fisher, E. M. C. Localization of genes encoding two human one-domain members of the AAA family: PSMC5 (the thyroid hormone receptor-interacting protein, TRIP1) and PSMC3 (the Tat-binding protein, TBP1). Hum. Genet. 99: 285-288, 1997

NCBI and Uniprot Product Information

• NCBI GI #: 21361144
• NCBI GeneID: 5702
• NCBI Accession #: NP_002795.2
• NCBI GenBank Nucleotide #: NM_002804.4
• UniProt Accession #: P17980; Q3B757; Q3B865; Q53HU5; Q6GPG8; Q6IBS1; Q96HD3; B2R8V1
• Molecular Weight: 49,204 Da
• NCBI Official Full Name: 26S protease regulatory subunit 6A
• NCBI Official Synonym Full Names: proteasome (prosome, macropain) 26S subunit, ATPase, 3
• NCBI Official Symbol: PSMC3
• NCBI Official Synonym Symbols: TBP1
• NCBI Protein Information: 26S protease regulatory subunit 6A; TBP-1; Tat-binding protein 1; proteasome subunit P50; proteasome 26S subunit ATPase 3; 26S proteasome AAA-ATPase subunit RPT5; human immunodeficiency virus tat transactivator binding protein-1
• UniProt Protein Name: 26S protease regulatory subunit 6A
• UniProt Gene Name: PSMC3
• UniProt Synonym Gene Names: TBP1; TBP-1
• UniProt Entry Name: PRS6A_HUMAN