Proteasome 20S beta 6 Rabbit Polyclonal Antibody
cat.: HA500349
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Polyclonal
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: Immunogen affinity purified.
Molecular weight: Predicted band size: 25 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Proteasome 20S beta 6 aa 21-239 / 239.
Positive control: C2C12 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Hela cell lysate, Jurkat cell lysate, Mouse colon tissue lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, HeLa, C6, NIH/3T3.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:500-1:5,000
1:200
1:500-1:1,000
Uniprot #: SwissProt: P28072 Human | Q60692 Mouse | P28073 Rat
Alternative names: DELTA LMPY Macropain delta chain MGC5169 Multicatalytic endopeptidase complex delta chain Proteasome (prosome macropain) subunit beta type 6 Proteasome beta 6 subunit Proteasome catalytic subunit 1 Proteasome delta chain Proteasome subunit beta 6 Proteasome subunit beta type 6 Proteasome subunit beta type-6 Proteasome subunit delta Proteasome subunit Y PSB6_HUMAN PSMB6 PSY large multifunctional protease Y Y
Images
HA500349_1.jpg Fig1: Western blot analysis of Proteasome 20S beta 6 on different lysates with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) at 1/5,000 dilution.

Lane 1: C2C12 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 6 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 (HA500349) at 1/5,000 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.
HA500349_2.jpg Fig2: Western blot analysis of Proteasome 20S beta 6 on different lysates with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) at 1/500 dilution.

Lane 1: Hela cell lysate (10 µg/Lane)
Lane 2: Jurkat cell lysate (10 µg/Lane)
Lane 3: Mouse colon tissue lysate (20 µg/Lane)
Lane 4: Mouse kidney tissue lysate (20 µg/Lane)
Lane 5: Rat kidney tissue lysate (20 µg/Lane)

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

Exposure time: 1 minute;

12% 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 (HA500349) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HA500349_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling Proteasome 20S beta 6 with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) 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-Proteasome 20S beta 6 antibody (HA500349) 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.
HA500349_4.jpg Fig4: Immunocytochemistry analysis of C6 cells labeling Proteasome 20S beta 6 with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) 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-Proteasome 20S beta 6 antibody (HA500349) 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.
HA500349_5.jpg Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling Proteasome 20S beta 6 with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Proteasome 20S beta 6 antibody (HA500349) at 1/200 dilution in 2% negative goat serum 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.
HA500349_6.jpg Fig6: Flow cytometric analysis of HeLa cells labeling Proteasome 20S beta 6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500349, 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).
HA500349_7.jpg Fig7: Flow cytometric analysis of NIH/3T3 cells labeling Proteasome 20S beta 6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500349, 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).
HA500349_8.jpg Fig8: Flow cytometric analysis of C6 cells labeling Proteasome 20S beta 6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500349, 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).
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