重组Anti-PBR抗体[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
Related conjugates and formulations
概述
-
产品名称
Anti-PBR抗体[EPR5384]
参阅全部 PBR 一抗 -
描述
兔单克隆抗体[EPR5384] to PBR -
宿主
Rabbit -
特异性
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.
-
经测试应用
适用于: IHC-Fr, Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab170987) -
阳性对照
- 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.
-
常规说明
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
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.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR5384 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab109497于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
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.
|
ICC/IF | (4) |
Use a concentration of 1 µg/ml.
|
说明 |
---|
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. |
ICC/IF
Use a concentration of 1 µg/ml. |
靶标
-
功能
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. -
组织特异性
Found in many tissue types. Expressed at the highest levels under normal conditions in tissues that synthesize steroids. -
序列相似性
Belongs to the TspO/BZRP family. -
细胞定位
Mitochondrion membrane. - Information by UniProt
-
数据库链接
- 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
-
别名
- Benzodiazapine receptor (peripheral) antibody
- Benzodiazepine peripheral binding site antibody
- BPBS antibody
see all
图片
-
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) 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.
-
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) 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.
-
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 kDaLanes 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.
-
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) 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.
-
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).
-
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.
-
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) 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)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)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.
-
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 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).
-
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.
-
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.
-
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) 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.
-
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.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (97)
ab109497 被引用在 97 文献中.
- Guilarte TR et al. Imaging neuroinflammation with TSPO: A new perspective on the cellular sources and subcellular localization. Pharmacol Ther 234:108048 (2022). PubMed: 34848203
- Foray C et al. Interrogating Glioma-Associated Microglia and Macrophage Dynamics Under CSF-1R Therapy with Multitracer In Vivo PET/MRI. J Nucl Med 63:1386-1393 (2022). PubMed: 35115369
- Varlow C et al. Characterization of neuroinflammatory positron emission tomography biomarkers in chronic traumatic encephalopathy. Brain Commun 4:fcac019 (2022). PubMed: 35198978
- Zhang X et al. TSPO Deficiency Exacerbates GSDMD-Mediated Macrophage Pyroptosis in Inflammatory Bowel Disease. Cells 11:N/A (2022). PubMed: 35269479
- Wang J et al. Inhibition of translocator protein 18 kDa suppressed the progression of glioma via the ELAV-like RNA-binding protein 1/MAPK-activated protein kinase 3 axis. Bioengineered 13:7457-7470 (2022). PubMed: 35285415