WB result of Rab5 Recombinant Rabbit mAb
Primary antibody: Rab5 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: MCF7 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 24 kDa
Observed MW: 24 kDa
Rab5 Recombinant Rabbit mAb (S-1677-74)
Rab5 Recombinant Rabbit mAb (S-1677-74)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Rab5 |
| Synonyms | Ras-related protein Rab-5A; RAB5A |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Cell membrane, Endosome |
| Accession | P20339 |
| Clone Number | S-1677-74 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | MCF7, HeLa, NIH/3T3, PC-12 |
| 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 |
| IP | 1:50 | Hu |
| ICC | 1:100 | Hu, Ms |
Background
Rab5 is a key regulator of the early endosome compartment and plays a crucial role in the control of endocytosis and membrane trafficking within the cell. It is one of the primary determinants of early endosomes, functioning as an important modulator of endosome progression and maturation. Rab5 orchestrates the fusion of endocytic vesicles with early endosomes and is involved in the recruitment of various effector proteins that facilitate vesicle docking, fusion, and the regulation of endosome motility along the cytoskeleton. The transition from early to late endosomes is regulated by a switch from Rab5 to Rab7, where Rab5 is gradually replaced by Rab7 on the same endosome. This transition is critical for the sorting of internalized cargo and the maturation of endosomes into multivesicular bodies. Rab5's activity is also associated with the regulation of hematopoietic homeostasis, as its inhibition can lead to cell overproliferation and disruptions in blood cell progenitor maintenance, highlighting its importance in cellular signaling pathways.
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Western Blot
WB result of Rab5 Recombinant Rabbit mAb
Primary antibody: Rab5 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 24 kDa
Observed MW: 24 kDa
WB result of Rab5 Recombinant Rabbit mAb
Primary antibody: Rab5 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 24 kDa
Observed MW: 24 kDa
IP
Rab5 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Rab5 in 0.4 mg MCF7 whole cell lysate.
Western blot was performed on the immunoprecipitate using Rab5 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: MCF7 whole cell lysate 20 µg (Input)
Lane 2: Rab5 Rabbit mAb IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in MCF7 whole cell lysate
Predicted MW: 24 kDa
Observed MW: 24 kDa
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Rab5 antibody was used at 1/100 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-Rab5 antibody was used at 1/100 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).
