WB result of UBE2F Recombinant Rabbit mAb
Primary antibody: UBE2F Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Jurkat whole cell lysate 20 µg
Lane 2: A375 whole cell lysate 20 µg
Lane 3: HeLa whole cell lysate 20 µg
Lane 4: HL-60 whole cell lysate 20 µg
Lane 5: NTERA2 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 21 kDa
Observed MW: 20 kDa
UBE2F Recombinant Rabbit mAb (S-2672-26)
UBE2F Recombinant Rabbit mAb (S-2672-26)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | UBE2F |
| Synonyms | NEDD8-conjugating enzyme UBE2F; NEDD8 carrier protein UBE2F; NEDD8 protein ligase UBE2F; NEDD8-conjugating enzyme 2; RING-type E3 NEDD8 transferase UBE2F; Ubiquitin-conjugating enzyme E2 F; NCE2 |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | Q969M7 |
| Clone Number | S-2672-26 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | Jurkat, A375, HeLa, HL-60, NTERA2, RAW264.7, NIH/3T3, mouse testis, rat ovary, rat testis |
| Predicted Reactivity | Bv |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt |
| IHC-P | 1:1000 | Hu, Ms, Rt |
| ICC | 1:500 | Hu, Ms |
| ICFCM | 1:50 | Hu, Ms |
Background
UBE2F (ubiquitin-conjugating enzyme E2 F) is a NEDD8-specific E2 conjugating enzyme that, following activation by the NAE1–UBA3 heterodimer, transfers NEDD8 onto CRL5–RBX2/SAG complexes to catalyze CUL5 neddylation, thereby activating CRL5 E3 ligase activity and promoting K11-linked polyubiquitylation and proteasomal degradation of the pro-apoptotic protein NOXA, a pathway that is frequently up-regulated in non-small-cell lung cancer to suppress apoptosis and drive tumor survival, and whose oncogenic action can be pharmacologically targeted to enhance chemotherapy sensitivity.
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Western Blot
WB result of UBE2F Recombinant Rabbit mAb
Primary antibody: UBE2F Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: RAW264.7 whole cell lysate 20 µg
Lane 2: NIH/3T3 whole cell lysate 20 µg
Lane 3: mouse testis lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 21 kDa
Observed MW: 20 kDa
WB result of UBE2F Recombinant Rabbit mAb
Primary antibody: UBE2F Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: rat testis lysate 20 µg
Lane 2: rat ovary lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 21 kDa
Observed MW: 20 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) labelling UBE2F antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) labelling UBE2F antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human testis. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human ovarian cancer. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human thyroid cancer. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse testis. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-UBE2F antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in Jurkat cells. Anti-UBE2F antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ICC shows positive staining in NIH/3T3 cells. Anti-UBE2F antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
