Validated using a knockout cell line
Recombinant
RabMAb

Anti-PBR antibody [EPR5384] (ab109497)

Overview

  • Product name
    Anti-PBR antibody [EPR5384]
    See all PBR primary antibodies
  • Description
    Rabbit monoclonal [EPR5384] to PBR
  • Specificity
    IHC results on rat tissues (such as liver and kidney) showed weak cytoplasmic and nuclear staining. However, other customer feedback suggests that this antibody works well in rat. Due to the inconclusive nature of these results, we do not currently guarantee this antibody in rat. Please contact our Scientific support team for more information.
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PBR aa 150 to the C-terminus (C terminal).
    (Peptide available as ab170987)

  • Positive control
    • WB: U-87MG, 293T, A431, RAW264.7, and NIH3T3 cell lysates IHC-P: Human bladder carcinoma and colon tissue ICC/IF: A431, U-87MG cells, HeLa cells, TSPO-HAP1 cells Flow Cyt: HepG2 and U-87MG cells. IP: A431 and U-87MG cell lysate.
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:

    Anti-PBR antibody (Alexa Fluor® 488) [EPR5384] (ab199779)

    Anti-PBR antibody (Alexa Fluor® 647) [EPR5384] (ab199836)

    Anti-PBR antibody (HRP) [EPR5384] (ab199548)

    Anti-PBR antibody (Alexa Fluor® 555) [EPR5384] (ab208060)

    Anti-PBR antibody (Alexa Fluor® 568) [EPR5384] (ab208061)

    Anti-PBR antibody (Phycoerythrin) [EPR5384] (ab208836)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Applications

Our Abpromise guarantee covers the use of ab109497 in the following tested applications.

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

Application Abreviews Notes
WB 1/10000 - 1/50000. Predicted molecular weight: 19 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/50 - 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Responsible for the manifestation of peripheral-type benzodiazepine recognition sites and is most likely to comprise binding domains for benzodiazepines and isoquinoline carboxamides. May play a role in the transport of porphyrins and heme. Plays a role in the transport of cholesterol across mitochondrial membranes in steroidogenic cells.
  • Tissue specificity
    Found in many tissue types. Expressed at the highest levels under normal conditions in tissues that synthesize steroids.
  • Sequence similarities
    Belongs to the TspO/BZRP family.
  • Cellular localization
    Mitochondrion membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Benzodiazapine receptor (peripheral) antibody
    • Benzodiazepine peripheral binding site antibody
    • BPBS antibody
    • BZRP antibody
    • DBI antibody
    • IBP antibody
    • Isoquinoline carboxamide-binding protein antibody
    • MBR antibody
    • mDRC antibody
    • Mitochondrial benzodiazepine receptor antibody
    • PBR antibody
    • PBS antibody
    • Peripheral benzodiazepine receptor antibody
    • Peripheral benzodiazepine receptor-related protein antibody
    • Peripheral type benzodiazepine receptor antibody
    • Peripheral-type benzodiazepine receptor antibody
    • pk18 antibody
    • PKBS antibody
    • PTBR antibody
    • Ptbzr antibody
    • PTBZR02 antibody
    • RATPTBZR02 antibody
    • translocator protein (18kDa) antibody
    • Translocator protein antibody
    • Tspo antibody
    • Tspo1 antibody
    • TSPOA_HUMAN antibody
    see all

Anti-PBR antibody [EPR5384] images



  • Predicted band size : 19 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PBR knockout HAP1 cell lysate (20 µg)
    Lane 3: HEK293 cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109497 observed at 15 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab109497 was shown to specifically react with PBR when PBR knockout samples were used. Wild-type and PBR knockout samples were subjected to SDS-PAGE.  Ab109497 and ab8245 (loading control to GAPDH) were diluted at 1/10000 and 1/10000 dilution respectively and incubated overnight at 4C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab109497 staining PBR in wild-type HAP1 cells (top panel) and PBR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109497 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab109497 staining PBR in HeLa. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109497 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling PBR with purified ab109497 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling PBR with purified ab109497 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • All lanes : Anti-PBR antibody [EPR5384] (ab109497) at 1/50000 dilution (purified)

    Lane 1 : RAW264.7 cell lysate
    Lane 2 : NIH/3T3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 19 kDa
    Observed band size : 18 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PBR antibody [EPR5384] (ab109497) at 1/50000 dilution (purified)

    Lane 1 : U87-MG cell lysate
    Lane 2 : HEK293 cell lysate
    Lane 3 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 19 kDa
    Observed band size : 18 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of U87-MG cells labelling PBR with purified ab109497 at 1/150 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ab109497 (purified) at 1/60 immunoprecipitating PBR in U87-MG whole cell lysate.

    Lane 1 (input): U87-MG whole cell lysate (10µg)

    Lane 2 (+): ab109497 + U87-MG whole cell lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109497 in U87-MG whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • ab109497 (purified) at 1/60 immunoprecipitating PBR in A431 whole cell lysate.

    Lane 1 (input): A431 whole cell lysate (10µg)

    Lane 2 (+): ab109497 + A431 whole cell lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109497 in A431 whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PBR antibody [EPR5384] (ab109497) at 1/10000 dilution (unpurified)

    Lane 1 : U-87 MG cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : RAW264.7 cell lysate
    Lane 5 : NIH3T3 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 19 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling PBR with unpurified ab109497 at a dilution of 1/100.

  • Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling PBR with unpurified ab109497 at a dilution of 1/50.

  • Overlay histogram showing HepG2 cells stained with unpurified ab109497 (red line). The cells were fixed with 4% paraformaldehyde (10 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. The cells were then incubated with the antibody (unpurified ab109497, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-PBR antibody [EPR5384] (ab109497)

This product has been referenced in:
  • Song D  et al. Complement C5a receptor knockout has diminished light-induced microglia/macrophage retinal migration. Mol Vis 23:210-218 (2017). IHC-Fr ; Mouse . Read more (PubMed: 28442885) »
  • Li HD  et al. A translocator protein 18 kDa agonist protects against cerebral ischemia/reperfusion injury. J Neuroinflammation 14:151 (2017). Read more (PubMed: 28754131) »

See all 17 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Muscle)
Gel Running Conditions
Reduced Denaturing (15% Acrylamide gel)
Loading amount
40 µg
Specification
Muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Ms. Lydie Combaret

Verified customer

Submitted Jul 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - other (Muscle)
Gel Running Conditions
Reduced Denaturing (15 Acrylamide Gel)
Loading amount
25 µg
Specification
Muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Ms. Lydie Combaret

Verified customer

Submitted Jul 10 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole mouse brain)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Gly Gel)
Loading amount
20 µg
Specification
Whole mouse brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jun 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Cell (mouse astrocytes)
Specification
mouse astrocytes
Permeabilization
Yes - 0.5% TX100
Fixative
Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Jan 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (Fibroblasts)
Specification
Fibroblasts
Permeabilization
Yes - 0.5% TX100
Fixative
Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Dec 12 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15)
Sample
Human Cell lysate - whole cell (HEK-293T cell line)
Specification
HEK-293T cell line
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 19 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Mouse Cell lysate - whole cell (BV2 cell line)
Specification
BV2 cell line
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 19 2014

Thank you for contacting us.

This lot has been sold out whihc had concentration 0.7340 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us.

We have 2 lots of this product, one has conc. of XXXmg/ml and other XXXmg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (prostate)
Specification
prostate
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 100mM sodium cytrate
Permeabilization
No
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Ms. Hadas Pahima

Verified customer

Submitted Oct 10 2012

1-10 of 12 Abreviews or Q&A

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