PSMB5 / MB1 Recombinant Rabbit Monoclonal Antibody [PSH15-20]
cat.: HA723724
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: PSH15-20
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 28 kDa
Isotype: IgG
Immunogen: Recombinant protein within human PSMB5 aa 34-263 / 263.
Positive control: HeLa cell lysate, MCF7 cell lysate, K-562 cell lysate, HEK-293 cell lysate, COS-1 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, HeLa, PC-12, human breast cancer tissue, human kidney tissue, human liver tissue, mouse kidney tissue, mouse liver tissue, rat kidney tissue, rat liver tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:10,000
1:100-1:200
1:2,000
1:1,000
Uniprot #: SwissProt: P28074 Human | O55234 Mouse | P28075 Rat
Alternative names: DKFZp459C139 EC 3.4.25.1 LMPX Macropain epsilon chain MB1 MGC104214 MGC118075 MGC134464 Multicatalytic endopeptidase complex epsilon chain Proteasome (prosome, macropain) subunit, beta type, 5 Proteasome beta 5 subunit Proteasome catalytic subunit 3 Proteasome chain 6 Proteasome epsilon chain Proteasome subunit beta type-5 Proteasome subunit MB1 Proteasome subunit X Proteasome subunit, beta type, 5 Proteasome subunit, beta-5 PSB5_HUMAN PSMB5 PSX large multifunctional protease X X
Images
HA723724_1.jpg Fig1: Western blot analysis of PSMB5 / MB1 on different lysates with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/10,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)
Lane 4: HEK-293 cell lysate (20 µg/Lane)
Lane 5: COS-1 cell lysate (20 µg/Lane)
Lane 6: Neuro-2a cell lysate (20 µg/Lane)
Lane 7: NIH/3T3 cell lysate (20 µg/Lane)
Lane 8: PC-12 cell lysate (20 µg/Lane)
Lane 9: Mouse kidney tissue lysate (40 µg/Lane)
Lane 10: Rat kidney tissue lysate (40 µg/Lane)

Predicted band size: 28 kDa
Observed band size: 28/22 kDa

Exposure time: 25 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723724) at 1/10,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723724_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PSMB5 / MB1 with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723724_3.jpg Fig3: Immunocytochemistry analysis of PC-12 cells labeling PSMB5 / MB1 with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723724_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723724) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723724_11.jpg Fig11: Flow cytometric analysis of HeLa cells labeling PSMB5 / MB1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723724, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA723724_12.jpg Fig12: Flow cytometric analysis of PC-12 cells labeling PSMB5 / MB1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723724, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA723724_13.jpg Fig13: Western blot analysis of PSMB5 / MB1 on different lysates with Rabbit anti-PSMB5 / MB1 antibody (HA723724) at 1/10,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 100ng/mL IFN gamma for 72 hours cell lysate
Lane 3: HeLa cell lysate
Lane 4: HeLa treated with 10ng/mL IFN alpha 1 for 16 hours cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 28 kDa
Observed band size: 22 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723724) at 1/10,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.