ET7107-36

Proteasome 20S LMP7 Recombinant Rabbit Monoclonal Antibody [JB54-32]

cat.: ET7107-36

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JB54-32

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 30 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human Proteasome 20S LMP7 aa 160-276 / 276.
Positive control:	U-937 cell lysate, A431 cell lysate, U-937, HUVEC, human kidney tissue, mouse kidney tissue, mouse small intestine tissue, rat kidney tissue.
Subcellular location:	Nucleus. Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:1,000 1:100 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: P28062 Human \| P28063 Mouse \| P28064 Rat
Alternative names:	ALDD D6S216 D6S216E Large multifunctional peptidase 7 Large multifunctional protease 7 LMP 7 LMP7 Low molecular mass protein 7 Low molecular weight protein 7 Macropain subunit C13 MGC1491 Multicatalytic endopeptidase complex subunit C13 NKJO OTTHUMP00000062981 Protease component C13 Proteasome (prosome macropain) subunit beta type 8 Proteasome (prosome, macropain) subunit, beta type, 8 (large multifunctional peptidase 7) Proteasome beta 8 subunit Proteasome catalytic subunit 3i Proteasome component C13 Proteasome related gene 7 Proteasome subunit beta 5i Proteasome subunit beta 8 Proteasome subunit beta type 8 Proteasome subunit beta type Proteasome subunit beta type-8 Proteasome subunit beta-5i Proteasome subunit Y2 PSB8_HUMAN PSMB 8 PSMB5i PSMB8 Really interesting new gene 10 protein RING 10 RING10 Y2

Images

	Fig1: Western blot analysis of Proteasome 20S LMP7 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-36, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: U-937 cell lysate Lane 2: A431 cell lysate
	Fig2: Immunocytochemistry analysis of U-937 cells labeling Proteasome 20S LMP7 with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) 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 LMP7 antibody (ET7107-36) 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/1,000 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 (ET7107-36) at 1/1,000 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.

	Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/200 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 (ET7107-36) at 1/200 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 small intestine tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/1,000 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 (ET7107-36) at 1/1,000 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 kidney tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/200 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 (ET7107-36) at 1/200 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 U-937 cells labeling Proteasome 20S LMP7.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-36, 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).

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