K27-linkage Specific Ubiquitin Recombinant Rabbit Monoclonal Antibody [JE75-30]
Catalog# HA723066
K27-linkage Specific Ubiquitin Recombinant Rabbit Monoclonal Antibody [JE75-30]
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WB
-
IHC-P
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IF-Cell
-
FC
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Dot Blot
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
K27-linkage Specific Ubiquitin Recombinant Rabbit Monoclonal Antibody [JE75-30]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Ubiquitin aa 20-34 (P0CG48).
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, FC, Dot Blot
Molecular Weight
Predicted band size: 26 kDa
Positive Control
Jurkat, human spleen tissue, mouse spleen tissue, rat spleen tissue.
Conjugation
unconjugated
Clone Number
JE75-30
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000
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IHC-P
-
1:200-1:1,000
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IF-Cell
-
1:1,000
-
FC
-
1:1,000
-
Dot Blot
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1:1,000
Target
Function
Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.
Background References
暂无
Subcellular Location
Cell Membrane, Cytoplasmic and Nuclear
UNIPROT
Synonyms
Epididymis secretory protein Li 50 antibody
FLJ25987 antibody
HEL S 50 antibody
MGC8385 antibody
Polyubiquitin B antibody
RPS 27A antibody
RPS27A antibody
UBA 52 antibody
UBA 80 antibody
UBA52 antibody
ExpandImages
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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. -
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. -
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. -
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. -
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). -
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"