Anti-PBR 抗体 [EPR5384] - BSA and Azide free (ab213654)
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 - BSA and Azide free
- Suitable for: IHC-Fr, IHC-P, WB, IP, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-PBR antibody [EPR5384] - BSA and Azide free
PBR 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR5384] to PBR - BSA and Azide free -
由来種
Rabbit -
特異性
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.
-
アプリケーション
適用あり: IHC-Fr, IHC-P, WB, IP, ICC/IF, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab170987) -
ポジティブ・コントロール
- WB: HCT116, HAP1, HeLa, U-87MG, 293T, A431, RAW264.7, and NIH3T3 cell lysates; IHC-P: Human bladder carcinoma and colon tissue; ICC/IF: U-87MG cells, HeLa cells, TSPO-HAP1 cells; Flow Cyt (intra): HepG2 and U-87MG cells; IP: A431 and U-87MG cell lysate; IHC-Fr: Mouse kidney and adrenal gland tissue; IHC-P: Human bladder carcinoma and Mouse hypothalamus tissue.
-
特記事項
ab213654 is the carrier-free version of ab109497.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR5384 -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
- Anti-PBR antibody [EPR5384] (ab109497)
- Alexa Fluor® 488 Anti-PBR antibody [EPR5384] (ab199779)
- Alexa Fluor® 647 Anti-PBR antibody [EPR5384] (ab199836)
- PE Anti-PBR antibody [EPR5384] (ab208836)
- Alexa Fluor® 555 Anti-PBR antibody [EPR5384] (ab283733)
- Alexa Fluor® 594 Anti-PBR antibody [EPR5384] (ab303575)
- Alexa Fluor® 568 Anti-PBR antibody [EPR5384] (ab314401)
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab213654の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
IHC-Fr |
Use at an assay dependent concentration.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
WB |
Use at an assay dependent concentration. Predicted molecular weight: 19 kDa.
|
|
IP |
Use at an assay dependent concentration.
|
|
ICC/IF |
Use at an assay dependent concentration.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
---|
IHC-Fr
Use at an assay dependent concentration. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 19 kDa. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
-
機能
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. -
組織特異性
Found in many tissue types. Expressed at the highest levels under normal conditions in tissues that synthesize steroids. -
配列類似性
Belongs to the TspO/BZRP family. -
細胞内局在
Mitochondrion membrane. - Information by UniProt
-
参照データベース
- 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
-
別名
- Benzodiazapine receptor (peripheral) antibody
- Benzodiazepine peripheral binding site antibody
- BPBS antibody
see all
画像
-
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?This data was developed using the same antibody clone in a different buffer formulation (ab109497).
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) 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 kDaThis data was produced with ab109497, the same antibody in a diffrent forulation with BSA and Azide.
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.
-
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?This data was developed using the same antibody clone in a different buffer formulation (ab109497).
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) analysis of mouse hypothalamus tissue labelling PBR with ab109497 at 1/1000 dilution (1.03 mg/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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
-
Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
-
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)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
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 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
-
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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
-
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, 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling PBR with unpurified ab109497 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109497).
プロトコール
データシートおよび資料
-
Datasheet download
Certificate of Compliance
参考文献 (0)
ab213654 は論文での使用が確認できていません。