Western blot analysis of extracts from various cell types using Rab4 Antibody.
Confocal immunofluorescent analysis of HeLa cells using Rab5A (E6N8S) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of SK-MEL-28 cells using Rab7 (D95F2) XP® Rabbit mAb (green). Actin filaments were labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal Immunofluorescent analysis of HeLa cells using Rab9A (D52G8) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of A549 cells using Rab11 (D4F5) XP® Rabbit mAb (green). Actin filaments were labeled wtih DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using Rab5A (E6N8S) Mouse mAb.
Western blot analysis of extracts from various cell lines using Rab7 (D95F2) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using Rab9A (D52G8) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using Rab11 (D4F5) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Rab5A (E6N8S) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Rab5A (E6N8S) Mouse mAb (left) compared to concentration matched Mouse (G3A1) IgG1 Isotype Control #5415 (right).
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Rab5A (E6N8S) Mouse mAb.
Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, lane 3 is Rab5A (E6N8S) Mouse mAb without HeLa cell extracts, and lane 4 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5A (E6N8S) Mouse mAb.
Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5 (C8B1) Rabbit mAb #3547.
Western blot analysis of extracts from various cell lines using Rab5A (E6N8S) Mouse mAb.
|Rab4 Antibody 2167||20 µl||
||H M R Hm Mk||25||Rabbit|
|Rab5A (E6N8S) Mouse mAb 46449||20 µl||
||H M R Mk||25||Mouse IgG1|
|Rab7 (D95F2) XP® Rabbit mAb 9367||20 µl||
||H M R Mk||23||Rabbit IgG|
|Rab9A (D52G8) XP® Rabbit mAb 5118||20 µl||
||H M R Mk||23||Rabbit IgG|
|Rab11 (D4F5) XP® Rabbit mAb 5589||20 µl||
||H M R Mk||25||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
|Anti-mouse IgG, HRP-linked Antibody 7076||100 µl||
The Rab Family Antibody Sampler Kit provides an economical means to evaluate the presence and status of Rab proteins in cells. This kit provides enough primary and secondary antibodies to perform two Western blot experiments per primary antibody.
Each antibody in the Rab Family Antibody Sampler Kit detects endogenous levels of its target protein. The Rab11 (D45F5) XP Rabbit mAb detects endogenous levels of total Rab11 protein, including isoforms Rab11a and Rab11b.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly190 of human Rab5A protein, Glu188 of human Rab7 protein, the carboxy terminus of human Rab9A protein and the amino terminus of human Rab11 protein. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp128 of human Rab4 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Rab family proteins are GTPases and members of the Ras superfamily of monomeric G proteins. These membrane-associated proteins are involved in many aspects of vesicle-mediated transport, taking part in the initial vesicle formation, transport of vesicles along the cytoskeleton, and eventual fusion of vesicle and target membranes. Rab4 is localized at early endosomes/recycling endosomes and functions as a key regulator for sorting/recycling of membrane and proteins (1,2). Both Rab4A and Rab4B isoforms are localized to similar cellular compartments and are believed to have similar functions (4). Rab4 interacts with several Rab4 effectors in a complex on a special endosome site to promote membrane/protein recycling (1,3). Rab5 is localized to the plasma membrane and early endosome and functions as a key regulator of vesicle trafficking during early endocytosis (1). The conformational change between Rab5-GTP and Rab5-GDP is essential for its biological function as a rate-limiting regulator at multiple steps during endocytosis (1,5). Similar to Rab4, Rab5 also interacts with specific Rab5 effectors on a specialized endosomal Rab domain to promote recycling between endosome and the plasma membrane (1,5,6). Both Rab7 and Rab9 are located in late endosomes but exert different functions. Rab7 associates with the RIPL effector protein to control membrane trafficking from early to late endosome and to lysosomes (7,8). Rab7 also helps to regulate growth receptor endocytic trafficking and degradation, and maturation of phagosome and autophagic vacuoles (8-11). Rab9 interacts with its effector proteins p40 and TIP47 (12,13) to promote the MPR (mannose 6-phosphate receptor)-associated lysosomal enzyme transport between late endosomes and the trans Golgi network (14,15). Rab11 (isoforms Rab11a and Rab11b) functions as a key regulator in the recycling of perinuclear, plasma membrane and Golgi compartment endosomes (16,17). Despite some overlap, distinct differences exist between Rab11a and Rab11b in both their cellular distribution and functional roles. Rab11a is ubiquitously expressed while Rab11b is found mainly in the heart and brain (18,19). Like other Rab proteins, Rab11 functions when associated with Rab11 family interacting proteins (FIPs). The three distinct classes of Rab11 FIPs all share a conserved carboxy-terminal Rab-binding domain that allows Rab-FIP protein interaction. When bound together, these proteins are thought to regulate membrane-associated protein sorting (20,21).
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