WB result of Keap1 Recombinant Rabbit mAb
Primary antibody: Keap1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NK-92 whole cell lysate 20 µg
Lane 2: MCF7 whole cell lysate 20 µg
Lane 3: HepG2 whole cell lysate 20 µg
Lane 4: Jurkat whole cell lysate 20 µg
Lane 5: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 70 kDa
Observed MW: 60 kDa
Keap1 Recombinant Rabbit mAb (S-1520-8)
Keap1 Recombinant Rabbit mAb (S-1520-8)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Keap1 |
| Synonyms | Kelch-like ECH-associated protein 1; Cytosolic inhibitor of Nrf2 (INrf2); INRF2; KIAA0132; KLHL19 |
| Immunogen | Recombinant Protein |
| Location | Cytoplasm, Nucleus |
| Accession | Q14145 |
| Clone Number | S-1520-8 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | NK-92, MCF7, HepG2, Jurkat, HeLa, mouse lung, mouse kidney |
| Predicted Reactivity | Gor, Pg |
| 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 |
| IHC-P | 1:500 | Hu, Ms, Rt |
| ICC | 1:500 | Hu |
Background
Keap1 (Kelch-like ECH-associated protein 1) is a critical intracellular sensor and regulator of the Nrf2/ARE (antioxidant response element) signaling pathway. Keap1 is an adaptor protein for the Cul3-based E3 ubiquitin ligase complex, responsible for targeting Nrf2 (nuclear factor erythroid 2-related factor 2) for ubiquitination and proteasomal degradation. It contains 27 cysteine residues that can be modified by oxidative stress, leading to the dissociation of Nrf2 and its subsequent stabilization and nuclear translocation. Under normal conditions, Keap1 binds to Nrf2, targeting it for degradation. However, in response to oxidative stress, certain cysteine residues in Keap1 are modified, inhibiting its ubiquitin ligase activity and leading to Nrf2 accumulation in the nucleus where it activates the transcription of antioxidant and detoxifying genes. Mutations in Keap1 or its interaction with Nrf2 have been implicated in various diseases, including cancer, where somatic mutations in cancer cells occur with high frequency in the DLGex and ETGE motifs of Nrf2, disrupting the Keap1-Nrf2 binding and leading to constitutive accumulation of Nrf2, which supports malignant growth.
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Western Blot
WB result of Keap1 Recombinant Rabbit mAb
Primary antibody: Keap1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: mouse lung lysate 20 µg
Lane 2: mouse kidney lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 70 kDa
Observed MW: 60 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-Keap1 antibody was used at 1/500 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-Keap1 antibody was used at 1/500 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 tonsil. Anti-Keap1 antibody was used at 1/500 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 lung cancer. Anti-Keap1 antibody was used at 1/500 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-Keap1 antibody was used at 1/500 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 kidney. Anti-Keap1 antibody was used at 1/500 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-Keap1 antibody was used at 1/500 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 HepG2 cells. Anti- Keap1 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).
