Antibodies, Proteins, Kits and Reagents for Life Science

Key features and details

Rabbit polyclonal to RALBP1
Suitable for: ICC/IF, WB
Reacts with: Human
Isotype: IgG

Product name

Anti-RALBP1 antibody
See all RALBP1 primary antibodies
Description

Rabbit polyclonal to RALBP1
Host species

Rabbit
Tested Applications & Species

Application Species
ICC/IF
Human

WB
Human

See all applications and species data
Immunogen

Synthetic peptide corresponding to Human RALBP1 aa 600 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as ab33445)
Positive control
- WB: Jurkat, HEK293, Hap1, HeLa and A431 cell lysates. ICC/IF: HEK293 cells.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS

Batches of this product that have a concentration
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Polyclonal
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
KO cell lines
- Human RALBP1 knockout HeLa cell line (ab265404)
KO cell lysates
- Human RALBP1 knockout HeLa cell lysate (ab258167)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab33446 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
ICC/IF	Human
WB	Human
All applications	Rat

Application	Abreviews	Notes
ICC/IF		Use a concentration of 1 µg/ml.
WB	(1)	Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 76 kDa).

Notes
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 76 kDa).

Function

Can 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 specificity

Expressed ubiquitously but at low levels. Shows a strong expression in the erythrocytes.
Sequence similarities

Contains 1 Rho-GAP domain.
Cellular localization

Membrane.
Target information above from: UniProt accession Q15311 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10928 Human
- Entrez Gene: 84014 Rat
- Omim: 605801 Human
- SwissProt: Q15311 Human
- SwissProt: Q62796 Rat
- Unigene: 528993 Human
- Unigene: 7107 Rat
Alternative names
- RLIP1 antibody
- 76 kDa Ral-interacting protein antibody
- 76-kDa Ral-interacting protein antibody
- Dinitrophenyl S-glutathione ATPase antibody
- DNP-SG ATPase antibody
- Ral-interacting protein 1 antibody
- Ral-interacting protein 1, 76-KD antibody
- RalA-binding protein 1 antibody
- RalBP1 antibody
- RBP1_HUMAN antibody
- RIP1 antibody
- RLIP1 antibody
- RLIP76 antibody
see all

Western blot - Anti-RALBP1 antibody (ab33446)

All lanes : Anti-RALBP1 antibody (ab33446) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RALBP1 knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 76 kDa

Lanes 1-4: Merged signal (red and green). Green - ab33446 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

ab33446 Anti-RALBP1 antibody 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. ab33446 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.
Western blot - Anti-RALBP1 antibody (ab33446)

All lanes : Anti-RALBP1 antibody (ab33446) at 1 µg/ml

Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 76 kDa
Observed band size: 90 kDa
why is the actual band size different from the predicted?

The band seen at 90 kDa is consistent with the banding pattern observed for other commercially available antibodies to RIP1.
Immunocytochemistry/ Immunofluorescence - Anti-RALBP1 antibody (ab33446)

ICC/IF image of ab33446 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33446, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

Western blot protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheet

Publishing research using ab33446? Please let us know so that we can cite the reference in this datasheet.

ab33446 has been referenced in 1 publication.

Johansson J et al. RAL GTPases Drive Intestinal Stem Cell Function and Regeneration through Internalization of WNT Signalosomes. Cell Stem Cell 24:592-607.e7 (2019). PubMed: 30853556

Western blot - Anti-RALBP1 antibody (ab33446)

All lanes : Anti-RALBP1 antibody (ab33446) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RALBP1 knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 76 kDa

Lanes 1-4: Merged signal (red and green). Green - ab33446 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

ab33446 Anti-RALBP1 antibody 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. ab33446 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.
Western blot - Anti-RALBP1 antibody (ab33446)

All lanes : Anti-RALBP1 antibody (ab33446) at 1 µg/ml

Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 76 kDa
Observed band size: 90 kDa
why is the actual band size different from the predicted?

The band seen at 90 kDa is consistent with the banding pattern observed for other commercially available antibodies to RIP1.
Immunocytochemistry/ Immunofluorescence - Anti-RALBP1 antibody (ab33446)

ICC/IF image of ab33446 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33446, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).