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ab69566 |
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Read our guarantee »Products:Signal Transduction >> Signaling Pathway >> G Protein Signaling >> Small G Proteins >> Ras Family
Anti-Rab25 antibody
See all Rab25 products (3) ...
Rabbit polyclonal to Rab25
ICC/IF, WBmore details
Reacts with
Rat, Human
Predicted to work with
Mouse, Rabbit, Horse, Dog, Chimpanzee
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Rab25.
(Peptide available as ab69566.)
This antibody gave a positive signal in the following Human Whole Cell lysates: MDA-MB-361, MCF7, T47D, HeLa, HepG2, Y79, SW480 and Jurkat This antibody also gave a positive signal in the following Tissue lysates: Human Breast Tumour and Rat Thymus
Liquid
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
Immunogen affinity purified
Polyclonal
IgG
Cancer >> Tumor biomarkers >> Oncoproteins
Signal Transduction >> Signaling Pathway >> G Protein Signaling >> Small G Proteins >> Ras Family
Our Abpromise guarantee covers the use of ab45855 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use a concentration of 5 µg/ml
WB: Use a concentration of 1 µg/mlDetects a band of approximately 32 kDa (predicted molecular weight: 23 kDa).
Involved in the regulation of cell survival. Promotes invasive migration of cells in which it functions to localize and maintain integrin alpha-V/beta-1 at the tips of extending pseudopodia. Increases the rates of ovarian and breast cancers progression and aggressiveness by modulating cellular processes such as proliferation, survival and migration of epithelial tumor cells. May selectively regulate the apical recycling pathway. Together with MYO5B regulates transcytosis.
Expressed in ovarian epithelium (NOE) and breast tissue. Expressed in ovarian cancer; expression is increased relative to NOE cells. Expression in ovarian cancer is stage dependent, with stage III and stage IV showing higher levels than early stage cancers. Expressed in breast cancer; expression is increased relative to normal breast tissue.
Belongs to the small GTPase superfamily. Rab family.
Cell membrane. Cell projection > pseudopodium membrane. Cytoplasmic vesicle. Colocalizes with integrin alpha-V/beta-1 in vesicles at the pseudopodial tips.
Target information above from: UniProt accessionP57735
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Western blot - Rab25 antibody (ab45855)

All lanes : Anti-Rab25 antibody (ab45855) at 1 µg/ml
Lane 1 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 3 : T47D (Human ductal breast epithelial tumor cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)
Additional bands at : 16 kDa,45 kDa,75 kDa. We are unsure as to the identity of these extra bands.
Western blot - Rab25 antibody (ab45855)

All lanes : Anti-Rab25 antibody (ab45855) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate (ab3957)
Lane 5 : Rat Thymus Tissue Lysate
Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 7 : Breast (Human) Tissue Lysate (ab30091)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 36 kDa (why is the actual band size different from the predicted?)
Additional bands at : 17 kDa,50 kDa,70 kDa. We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - Rab25 antibody (ab45855)

ICC/IF image of ab45855 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45855, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa and MCF7 cells at 5µg/ml.
ab45855 has not yet been referenced specifically in any publications.
Publishing research using ab45855? Please let us know so that we can cite the reference in this datasheet
Concentration of lot no. is
Concentration not available for this lot.
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All lanes : Anti-Rab25 antibody (ab45855) at 1 µg/ml
Lane 1 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 3 : T47D (Human ductal breast epithelial tumor cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)
Additional bands at : 16 kDa,45 kDa,75 kDa. We are unsure as to the identity of these extra bands.

All lanes : Anti-Rab25 antibody (ab45855) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate (ab3957)
Lane 5 : Rat Thymus Tissue Lysate
Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 7 : Breast (Human) Tissue Lysate (ab30091)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 36 kDa (why is the actual band size different from the predicted?)
Additional bands at : 17 kDa,50 kDa,70 kDa. We are unsure as to the identity of these extra bands.

ICC/IF image of ab45855 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45855, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa and MCF7 cells at 5µg/ml.
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