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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PBR antibody [EPR5384] (ab109497)

  • Datasheet
  • SDS
Reviews (14)Q&A (3)References (79)

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Western blot - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
  • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
  • Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
  • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
  • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
  • Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)
  • Anti-PBR antibody [EPR5384] (ab109497)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5384] to PBR
  • Suitable for: IHC-Fr, Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free PE

You may also be interested in

Primary
Product image
Alexa Fluor® 488 Anti-PBR antibody [EPR5384] (ab199779)
Knockout
Product image
Human TSPO (PBR) knockout HeLa cell line (ab264942)
Knockout
Product image
Human TSPO (PBR) knockout HCT116 cell line (ab266878)

View more associated products

Overview

  • Product name

    Anti-PBR antibody [EPR5384]
    See all PBR primary antibodies
  • Description

    Rabbit monoclonal [EPR5384] to PBR
  • Host species

    Rabbit
  • 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: IHC-Fr, Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab170987)

  • Positive control

    • WB: HCT116, HAP1, HeLa, U-87MG, 293T, A431, RAW264.7, and NIH3T3 cell lysates. Mouse lung, kidney, testis, brain, heart, liver and spleen tissue lysates. IHC-P: Human bladder carcinoma and colon tissue. Mouse lung, kidney, testis, brain, heart, liver and spleen tissue. IHC-Fr: Mouse kidney, adrenal gland tissue. ICC/IF: U-87MG, HeLa, MA-10 and TSPO-HAP1 cells; HeLa-TSPO (negative). Flow Cyt (intra): HepG2 and U-87MG cells. IP: A431 and U-87MG cell lysates.
  • General notes

    The Peripheral-Type Benzodiazepine Receptor (PBR) is a protein which is primarily found on mitochondria, mediating cholesterol transport across the mitochondrial membranes in steroidogenic cells. PBR has been shown to play a regulatory role in tumor cell proliferation.

    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
    For more information see here.

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

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5384
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calcium Channels
    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Metabolism
    • Types of disease
    • Cancer
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-PBR antibody [EPR5384] (ab199779)
    • Alexa Fluor® 647 Anti-PBR antibody [EPR5384] (ab199836)
    • PE Anti-PBR antibody [EPR5384] (ab208836)
    • Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human TSPO (PBR) knockout HeLa cell line (ab264942)
    • Human TSPO (PBR) knockout HCT116 cell line (ab266878)
  • KO cell lysates

    • Human TSPO (PBR) knockout HeLa cell lysate (ab257066)
    • Human TSPO (PBR) knockout HCT116 cell lysate (ab257067)
  • Positive Controls

    • A-431 membrane extract lysate (ab30134)
    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Pig PBR protein (Tagged) (ab235614)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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
IHC-Fr
1/250.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

Flow Cyt (Intra)
1/50 - 1/150.

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

WB (9)
1/10000 - 1/50000. Predicted molecular weight: 19 kDa.
IP
1/10 - 1/100.
IHC-P (1)
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF (4)
Use a concentration of 1 µg/ml.
Notes
IHC-Fr
1/250.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

Flow Cyt (Intra)
1/50 - 1/150.

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

WB
1/10000 - 1/50000. Predicted molecular weight: 19 kDa.
IP
1/10 - 1/100.
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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.
  • Target information above from: UniProt accession P30536 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 706 Human
    • Entrez Gene: 12257 Mouse
    • Omim: 109610 Human
    • SwissProt: B1AH88 Human
    • SwissProt: P30536 Human
    • SwissProt: P50637 Mouse
    • Unigene: 202 Human
    • Unigene: 1508 Mouse
    • 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

    Images

    • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      All lanes : Anti-PBR antibody [EPR5384] (ab109497) at 1/1000 dilution

      Lane 1 : Wild-type HCT116 cell lysate
      Lane 2 : TSPO knockout HCT116 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 17 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab109497 observed at 17 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109497 was shown to react with PBR in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266878 (knockout cell lysate ab257067) was used. Wild-type HCT116 and TSPO knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109497 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)

      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 - Anti-PBR antibody [EPR5384] (ab109497)
      Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)

      ab109497 staining PBR in wild-type HeLa cells (top panel) and TSPO knockout HeLa cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10min), 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 0.1 μg/ml and ab195884 at 2 μg/ml (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 (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)

      Immunohistochemistry (Frozen sections) analysis of mouse kidney tissue sections labeling PBR with Purified ab109497 at 1/250 (3.8 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

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

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

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 19 kDa



      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 in wild type HAP1 cells. No band was observed 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.

    • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      All lanes : Anti-PBR antibody [EPR5384] (ab109497) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : TSPO (PBR) knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 15 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab109497 observed at 15 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109497 was shown to react with PBR in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264942 (knockout cell lysate ab257066) was used. Wild-type HeLa and TSPO knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109497 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)

      Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) analysis of mouse hypothalamus tissue labelling PBR with ab109497 at 1/1000 dilution (1.03 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0 (ab93684). A goat anti-rabbit IgG (H+L) (HRP) was used as the secondary antibody (concentration: ready to use). A secondary-antibody-only control was also performed. Counterstained with hematoxylin.

      Positive staining was observed on ependymal cells and endothelial cells in mouse hypothalamus.

    • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
      Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)

      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).

    • Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)
      Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)

      Intracellular Flow Cytometry analysis of U-87MG 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. 

       

       

    • Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)
      Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)

      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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] (ab109497)

      Immunohistochemistry (Frozen sections) analysis of mouse adrenal gland tissue sections labeling PBR with Purified ab109497 at 1/250 (3.8 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)Wang, H. et al PLoS One. 2016 Dec 1;11(12):e0167307. doi: 10.1371/journal.pone.0167307. eCollection 2016 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      TSPO (PBR) expression was abolished in global KO mice without pathological changes

      TSPO expression in different tissues from WT and KO mice was detected by western blotting (A) and IHC (B). Scale Bars, 100μm.

      4% PFA-fixed tissue sections were blocked with 5% goat serum and incubated overnight at 4°C with ab109497 at 1/1500 dilution. DAB staining.

      Note: TSPO and PBR are alternative names for the same target.

      (Adapted from Figure 3 of Wang et al)

    • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
      Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)Morohaku, K. et al Send to PLoS One. 2013 Sep 5;8(9):e74509. doi: 10.1371/journal.pone.0074509. eCollection 2013 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      TSPO (PBR) expression is localized to the mitochondria

      (A) Panel shows confocal images of TSPO (red), VDAC1 (green) and nuclear counterstain (blue) in MA-10 Leydig (mouse). cells. (B) Negative control panel. Colocalization of TSPO to the mitochondrial protein VDAC1 validates the specific localization of TSPO. Scale bar 20 µm.

      Cells were fixed with 4% formaldehyde and permeabilized using 0.1% Triton X-100. Cells were then blocked using 5% normal goat serum and incubated with ab109497 at 1/200 dilution.

      Note: TSPO and PBR are alternative names for the same target.

    • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
      Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)

      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)
      Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] (ab109497)

      Immunocytochemistry/Immunofluorescence analysis of U-87MG 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).

    • Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      Western blot - Anti-PBR antibody [EPR5384] (ab109497)
      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
      All lanes : 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.

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

      Lane 1 : U-87MG cell lysate
      Lane 2 : HEK293 cell lysate
      Lane 3 : A431 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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.

    • Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)
      Immunoprecipitation - Anti-PBR antibody [EPR5384] (ab109497)

      ab109497 (purified) at 1/60 immunoprecipitating PBR in U-87MG whole cell lysate.

      Lane 1 (input): U-87MG whole cell lysate (10µg)

      Lane 2 (+): ab109497 + U-87MG whole cell lysate (10µg).

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

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (ab109497)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling PBR with unpurified ab109497 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)
      Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] (ab109497)

      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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. 

       

       

    • Anti-PBR antibody [EPR5384] (ab109497)
      Anti-PBR antibody [EPR5384] (ab109497)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (79)

    Publishing research using ab109497? Please let us know so that we can cite the reference in this datasheet.

    ab109497 has been referenced in 79 publications.

    • Barca C  et al. Impact of hydroxytyrosol on stroke: tracking therapy response on neuroinflammation and cerebrovascular parameters using PET-MR imaging and on functional outcomes. Theranostics 11:4030-4049 (2021). PubMed: 33754046
    • Thau-Zuchman O  et al. A new ketogenic formulation improves functional outcome and reduces tissue loss following traumatic brain injury in adult mice. Theranostics 11:346-360 (2021). PubMed: 33391479
    • Foray C  et al. Imaging temozolomide-induced changes in the myeloid glioma microenvironment. Theranostics 11:2020-2033 (2021). PubMed: 33500706
    • Zhang L  et al. Intravenous transplantation of olfactory ensheathing cells reduces neuroinflammation after spinal cord injury via interleukin-1 receptor antagonist. Theranostics 11:1147-1161 (2021). PubMed: 33391526
    • Tuominen S  et al. Evaluation of [18F]F-DPA as a target for TSPO in head and neck cancer under normal conditions and after radiotherapy. Eur J Nucl Med Mol Imaging 48:1312-1326 (2021). PubMed: 33340054
    View all Publications for this product

    Customer reviews and Q&As

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    1-10 of 17 Abreviews or Q&A

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Brain tissue (cerebellum))
    Gel Running Conditions
    Non-reduced Denaturing (12)
    Loading amount
    30 µg
    Specification
    Brain tissue (cerebellum)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 24 2022

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Dopaminergic Neurons)
    Gel Running Conditions
    Reduced Denaturing (NuPage 4-12% MES buffer)
    Loading amount
    30 µg
    Specification
    Dopaminergic Neurons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 22 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    African green monkey Cell (COS-7 Cell Line from African green monkey kidney)
    Permeabilization
    Yes - 0.5% TX100
    Specification
    COS-7 Cell Line from African green monkey kidney
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    HERR DR. Vladimir Milenkovic

    Verified customer

    Submitted Jan 16 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Gerbil Cell (Mongolian gerbil primary fibroblasts)
    Permeabilization
    Yes - 0.5% TX100
    Specification
    Mongolian gerbil primary fibroblasts
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    HERR DR. Vladimir Milenkovic

    Verified customer

    Submitted Oct 29 2019

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T, HCT116)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    35 µg
    Specification
    HEK293T, HCT116
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 28 2019

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Ms. Lydie Combaret

    Verified customer

    Submitted Jul 14 2017

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Ms. Lydie Combaret

    Verified customer

    Submitted Jul 10 2017

    Western blot abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 24 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    HERR DR. Vladimir Milenkovic

    Verified customer

    Submitted Jan 30 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PBR antibody [EPR5384]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    HERR DR. Vladimir Milenkovic

    Verified customer

    Submitted Dec 12 2014

    1-10 of 17 Abreviews or Q&A

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