ER31212

Ubiquitin Rabbit Polyclonal Antibody

cat.: ER31212

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

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 8 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human Ubiquitin aa 21-76.
Positive control:	293T cell lysate, HepG2 cell lysate, Hela cell lysate, MCF-7 cell lysate, Hela, HepG2, human tonsil tissue, mouse lung tissue, human kidney tissue.
Subcellular location:	Cytoplasm, nucleus, Membrane, Mitochondrion, Mitochondrion outer membrane.
Recommended Dilutions: WB IF-Cell IHC-P	1:500-1:1,000 1:100-1:200 1:200-1:1,000
Uniprot #:	SwissProt: P0CG48 Human
Alternative names:	Epididymis secretory protein Li 50 FLJ25987 HEL S 50 MGC8385 Polyubiquitin B RPS 27A RPS27A UBA 52 UBA 80 UBA52 UBA80 UBB UBB_HUMAN UBC UBCEP 1 UBCEP 2 UBCEP1 UBCEP2 Ubiquitin Ubiquitin B

Images

	Fig1: Western blot analysis of Ubiquitin on different lysates with Rabbit anti-Ubiquitin antibody (ER31212) at 1/500 dilution. Lane 1: MCF-7 cell lysate Lane 2: MCF-7 cell lysate treated with 50 µM MG132 for 1.5h Lysates/proteins at 10 µg/Lane. Predicted band size: 8 kDa Exposure time: 2 minutes; 15% 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 (ER31212) 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.
	Fig2: Western blot analysis of Ubiquitin on different cell lysates using anti-Ubiquitin antibody at 1/1,000 dilution. Positive control: Lane 1: 293T cell lysate Lane 2: HepG2 cell lysate Lane 3: Hela cell lysate Lane 4: MCF-7 cell lysate
	Fig3: ICC staining Ubiquitin in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

	Fig4: ICC staining Ubiquitin in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
	Fig5: ICC staining Ubiquitin in MCF-7 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Ubiquitin antibody (ER31212) 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 (ER31212) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Ubiquitin antibody (ER31212) 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 (ER31212) 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.

Fig8: Immunocytochemistry analysis of NIH/3T3 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 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-Ubiquitin antibody (ER31212) 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 (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.

Fig9: Immunocytochemistry analysis of PC-12 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 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-Ubiquitin antibody (ER31212) 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 (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.

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