Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RALBP1 antibody (ab33446)

Key features and details

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

Overview

  • 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.

Properties

Applications

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).

Target

  • 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.
  • Information by UniProt

  • Database links

    • 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

    Images

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

    References (1)

    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

    Images

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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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