K27-linkage Specific Ubiquitin Recombinant Rabbit Monoclonal Antibody [JE75-30]
cat.: HA723066
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC, Dot Blot |
| Clonality: | Monoclonal |
| Clone number: | JE75-30 |
| 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: | 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: 26 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human Ubiquitin aa 20-34 (P0CG48). |
| Positive control: | Jurkat, human spleen tissue, mouse spleen tissue, rat spleen tissue. |
| Subcellular location: | Cell Membrane, Cytoplasmic and Nuclear |
| Recommended Dilutions:
WB IHC-P IF-Cell FC Dot Blot |
1:1,000 1:200-1:1,000 1:1,000 1:1,000 1:1,000 |
| Uniprot #: | SwissProt: P0CG47 Human | P0CG48 Human | P62979 Human | P62987 Human | P62988 Human | P0CG49 Mouse | P62991 Mouse | P0CG51 Rat | P62989 Rat |
| 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
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Fig1:
Immunocytochemistry analysis of Jurkat cells labeling K27-linkage Specific Ubiquitin with Rabbit anti-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/1,000 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-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/1,000 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. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA723066) 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA723066) 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA723066) 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. |
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Fig5:
Flow cytometric analysis of Jurkat cells labeling K27-linkage Specific Ubiquitin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723066, 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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Fig6:
Dot blot analysis of K27-linkage Specific Ubiquitin on different proteins with Rabbit anti-K27-linkage Specific Ubiquitin antibody (HA723066) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Unmodified Ubiquitin peptide (negative) Lane 2: K27-linkage Specific Ubiquitin peptide (positive) Lane 3: K6-linkage Specific Ubiquitin peptide (negative) Lane 4: K11-linkage Specific Ubiquitin peptide (negative) Lane 5: K29-linkage Specific Ubiquitin peptide (negative) Lane 6: K33-linkage Specific Ubiquitin peptide (negative) Lane 5: K48-linkage Specific Ubiquitin peptide (negative) Lane 6: K63-linkage Specific Ubiquitin peptide (negative) Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 3 minutes; ECL: K1801. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.