HA500533

PSMB5 / MB1 Rabbit Polyclonal Antibody

cat.: HA500533

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 25 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human PSMB5 aa 34-263 / 263.
Positive control:	MCF7 cell lysate, PC-3M cell lysate, K-562 cell lysate, HepG2 cell lysate, LO2 cell lysate, HEK-293 cell lysate, A549 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, A431, HeLa, human breast carcinoma tissue, mouse liver tissue, rat brain tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:2,000 1:2,000 1:200 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

Fig1: Western blot analysis of PSMB5 / MB1 on different lysates with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/2,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: PC-3M cell lysate
Lane 3: K-562 cell lysate
Lane 4: HepG2 cell lysate
Lane 5: LO2 cell lysate
Lane 6: HEK-293 cell lysate
Lane 7: A549 cell lysate
Lane 8: NIH/3T3 cell lysate
Lane 9: Neuro-2a cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 28 kDa
Observed band size: 23/25 kDa

Exposure time: 45 seconds;

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 (HA500533) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of A431 cells labeling PSMB5 / MB1 with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (HA500533) 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 (M1305-2, 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.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling PSMB5 / MB1 with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 (HA500533) 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 (M1305-2, 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA500533) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA500533) 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PSMB5 / MB1 antibody (HA500533) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA500533) 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.
	Fig7: Flow cytometric analysis of A431 cells labeling PSMB5 / MB1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500533, 1μg/mL) (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).
	Fig8: Flow cytometric analysis of HeLa cells labeling PSMB5 / MB1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500533, 1μg/mL) (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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.