Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6472] to RALBP1
- Suitable for: WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-RALBP1 antibody [EPR6472]
See all RALBP1 primary antibodies
DescriptionRabbit monoclonal [EPR6472] to RALBP1
Tested applicationsSuitable for: WB, IHC-P, Flow Cytmore details
Unsuitable for: ICC/IF or IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to residues in Human RALBP1 (UniProt ID: Q15311).
- WB: Daudi, MCF7, HT-29, and HeLa cell lysates. IHC-P: Human lung tissue Flow Cyt: HEK-294T cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Dissociation constant (KD)KD = 5.42 x 10 -11 M Learn more about KD
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
KO cell lines
KO cell lysates
Our Abpromise guarantee covers the use of ab133549 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/50000. Detects a band of approximately 95 kDa (predicted molecular weight: 76 kDa).|
|IHC-P||1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionCan activate specifically hydrolysis of GTP bound to RAC1 and CDC42, but not RALA. Mediates ATP-dependent transport of S-(2,4-dinitrophenyl)-glutathione (DNP-SG) and doxorubicin (DOX) and is the major ATP-dependent transporter of glutathione conjugates of electrophiles (GS-E) and DOX in erythrocytes. Can catalyze transport of glutathione conjugates and xenobiotics, and may contribute to the multidrug resistance phenomenon. Serves as a scaffold protein that brings together proteins forming an endocytotic complex during interphase and also with CDK1 to switch off endocytosis, One of its substrates would be EPN1/Epsin.
Tissue specificityExpressed ubiquitously but at low levels. Shows a strong expression in the erythrocytes.
Sequence similaritiesContains 1 Rho-GAP domain.
- Information by UniProt
- RLIP1 antibody
- 76 kDa Ral-interacting protein antibody
- 76-kDa Ral-interacting protein antibody
All lanes : Anti-RALBP1 antibody [EPR6472] (ab133549) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RALBP1 knockout HeLa cell lysate
Lane 3 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 76 kDa
Lanes 1-3: Merged signal (red and green). Green - ab133549 observed at 95 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133549 Anti-RALBP1 antibody [EPR6472] was shown to specifically react with PDE10A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265404 (knockout cell lysate ab258167) was used. Wild-type and PDE10A knockout samples were subjected to SDS-PAGE. ab133549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-RALBP1 antibody [EPR6472] (ab133549) at 1/10000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : HT-29 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 76 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?
Overlay histogram showing HEK293 cells stained with ab133549 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133549, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunohistochemical analysis of paraffin embedded Human lung tissue labelling RALBP1 with ab133549 antibody at a dilution of 1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
ab133549 has been referenced in 4 publications.
- Wang W et al. RLIP76 increases apoptosis through Akt/mTOR signaling pathway in gastric cancer. Oncol Rep 36:2216-24 (2016). PubMed: 27572296
- Wang W et al. Downregulation of RLIP76 is associated with vincristine resistance in human colorectal cancer HCT-8/VCR cells. Int J Oncol 49:1505-1512 (2016). PubMed: 27633085
- Monot M et al. Early Steps of Jaagsiekte Sheep Retrovirus-Mediated Cell Transformation Involve the Interaction between Env and the RALBP1 Cellular Protein. J Virol 89:8462-73 (2015). PubMed: 26041289
- Kim JE et al. LIM kinase-2 induces programmed necrotic neuronal death via dysfunction of DRP1-mediated mitochondrial fission. Cell Death Differ : (2014). PubMed: 24561342