Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)

Rabbit Recombinant Monoclonal PBR antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, IHC-Fr, Flow Cyt (Intra) and reacts with Mouse, Human samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (AB213654), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	IHC-Fr	Flow Cyt (Intra)
Human	Expected	Tested	Tested	Tested	Expected	Tested
Mouse	Tested	Expected	Expected	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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6 products for Alternative Version

Target data

See full target information TSPO

Function

Can bind protoporphyrin IX and may play a role in the transport of porphyrins and heme (By similarity). Promotes the transport of cholesterol across mitochondrial membranes and may play a role in lipid metabolism (PubMed:24814875), but its precise physiological role is controversial. It is apparently not required for steroid hormone biosynthesis. Was initially identified as peripheral-type benzodiazepine receptor; can also bind isoquinoline carboxamides (PubMed:1847678).

Alternative names

BZRP, MBR, TSPO, Translocator protein, Mitochondrial benzodiazepine receptor, PKBS, Peripheral-type benzodiazepine receptor, PBR

Recommended products

Rabbit Recombinant Monoclonal PBR antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, IHC-Fr, Flow Cyt (Intra) and reacts with Mouse, Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR5384
Purification technique: Affinity purification Protein A
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.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab213654 is the carrier-free version of Anti-PBR antibody [EPR5384] ab109497.

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

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 batch production

For more information, read more on recombinant antibodies.

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

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PBR also known as the Peripheral Benzodiazepine Receptor is a protein predominantly found in the outer mitochondrial membrane. It has alternate names like Translocator Protein (TSPO) and the mass of this protein is approximately 18 kDa. PBR is expressed in various tissues but shows high levels in steroidogenic tissues like adrenal glands as well as in the brain heart liver and kidneys. The abundant presence in these tissues highlights its importance in a variety of physiological functions.

Biological function summary

PBR interacts with cholesterol for the synthesis of steroid hormones making it important for steroidogenesis. PBR is a part of the Mitochondrial Permeability Transition Pore complex (MPTP) involved in regulating the transport of molecules across the mitochondrial membrane. Through its association with the MPTP PBR plays a significant role in mitochondrial functions such as apoptosis and energy metabolism. The interaction with other molecules also includes the binding with benzodiazepines impacting processes like immune response and cell proliferation.

Pathways

PBR plays a role in the steroid biosynthesis and apoptosis pathways. It interfaces with the StAR (Steroidogenic Acute Regulatory) protein to facilitate cholesterol transport into mitochondria the initial step in steroid hormone production. PBR is also involved in pathways that regulate apoptosis and mitochondrial function linking it to different cellular processes through interactions with proteins like VDAC (Voltage-Dependent Anion Channel).

Associated diseases and disorders

PBR has relevance to conditions such as neurodegenerative diseases and cancer. PBR expression changes in disorders like Alzheimer's disease where it might reflect mitochondrial dysfunctions. It has also been associated with certain cancers where aberrant PBR activity might contribute to altered cell proliferation and apoptosis. Proteins like caspases are involved in the apoptotic pathways connected with PBR highlighting its involvement in disease processes.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

17 product images

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/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
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 Anti-PBR antibody [EPR5384] 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 (Anti-PBR antibody [EPR5384] ab109497).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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/
Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
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 Anti-PBR antibody [EPR5384] 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 (Anti-PBR antibody [EPR5384] ab109497).
Western blot - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
This data was developed using the same antibody clone in a different buffer formulation (Anti-PBR antibody [EPR5384] ab109497).
Lanes 1- 2: Merged signal (red and green). Green - Anti-PBR antibody [EPR5384] ab109497 observed at 17 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-PBR antibody [EPR5384] ab109497 was shown to react with PBR in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line Human TSPO (PBR) knockout HCT116 cell line ab266878 (knockout cell lysate Human TSPO (PBR) knockout HCT116 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. Anti-PBR antibody [EPR5384] ab109497 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.
All lanes: Western blot - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497) at 1/1000 dilution
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: TSPO knockout HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human TSPO (PBR) knockout HCT116 cell line (Human TSPO (PBR) knockout HCT116 cell line ab266878)
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 17 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling PBR with purified Anti-PBR antibody [EPR5384] ab109497 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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 (Anti-PBR antibody [EPR5384] ab109497).
Immunoprecipitation - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Anti-PBR antibody [EPR5384] ab109497 (purified) at 1/60 immunoprecipitating PBR in A431 whole cell lysate.
Lane 1 (input): A431 whole cell lysate (10μg)
Lane 2 (+): Anti-PBR antibody [EPR5384] ab109497 + A431 whole cell lysate (10μg).
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PBR antibody [EPR5384] ab109497 in A431 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (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 (Anti-PBR antibody [EPR5384] ab109497).
All lanes: Immunoprecipitation - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497)
Predicted band size: 19 kDa
Observed band size: 19 kDa
Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunohistochemistry (Frozen sections) analysis of mouse kidney tissue sections labeling PBR with Purified Anti-PBR antibody [EPR5384] 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, Goat Anti-Rabbit IgG H&L (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 (Anti-PBR antibody [EPR5384] ab109497).
Western blot - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
This data was produced with Anti-PBR antibody [EPR5384] ab109497, the same antibody in a diffrent forulation with BSA and Azide.
Lanes 1 - 4: Merged signal (red and green). Green - Anti-PBR antibody [EPR5384] ab109497 observed at 15 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-PBR antibody [EPR5384] 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. Anti-PBR antibody [EPR5384] ab109497 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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: Western blot - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497) at 1/10000 dilution
Lane 1: Wild-type HAP1 cell lysate at 20 µg
Lane 2: PBR knockout HAP1 cell lysate at 20 µg
Lane 3: HEK293 cell lysate at 20 µg
Lane 4: A431 cell lysate at 20 µg
Predicted band size: 19 kDa
Western blot - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
This data was developed using the same antibody clone in a different buffer formulation (Anti-PBR antibody [EPR5384] ab109497).
Lanes 1- 2: Merged signal (red and green). Green - Anti-PBR antibody [EPR5384] ab109497 observed at 15 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-PBR antibody [EPR5384] ab109497 was shown to react with PBR in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human TSPO (PBR) knockout HeLa cell line ab264942 (knockout cell lysate Human TSPO (PBR) knockout HeLa 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. Anti-PBR antibody [EPR5384] ab109497 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.
All lanes: Western blot - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: TSPO (PBR) knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human TSPO (PBR) knockout HeLa cell line (Human TSPO (PBR) knockout HeLa cell line ab264942)
Performed under reducing conditions.
Predicted band size: 112 kDa, 13 kDa, 134 kDa, 152 kDa, 17 kDa, 19 kDa, 25 kDa, 26 kDa, 28 kDa, 29 kDa, 37 kDa, 38 kDa, 39 kDa, 42 kDa, 43 kDa, 49 kDa, 50 kDa, 52 kDa, 56 kDa, 58 kDa, 68 kDa, 69 kDa, 71 kDa, 72 kDa, 74 kDa, 80 kDa, 81 kDa, 83 kDa, 87 kDa
Observed band size: 112 kDa, 15 kDa, 36 kDa, 38 kDa, 42 kDa, 45 kDa, 55 kDa, 60 kDa, 74 kDa, 80 kDa, 87 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) analysis of mouse hypothalamus tissue labelling PBR with Anti-PBR antibody [EPR5384] ab109497 at 1/1000 dilution (1.03 mg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9.0 (Antigen Retrieval Buffer (100X 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 (Anti-PBR antibody [EPR5384] ab109497).
Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Anti-PBR antibody [EPR5384] 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 Anti-PBR antibody [EPR5384] ab109497 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Anti-PBR antibody [EPR5384] ab109497).
Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Intracellular Flow Cytometry analysis of U87-MG cells labelling PBR with purified Anti-PBR antibody [EPR5384] 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 (Anti-PBR antibody [EPR5384] ab109497).
Immunohistochemistry (Frozen sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunohistochemistry (Frozen sections) analysis of mouse adrenal gland tissue sections labeling PBR with Purified Anti-PBR antibody [EPR5384] 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, Goat Anti-Rabbit IgG H&L (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 (Anti-PBR antibody [EPR5384] ab109497).
Immunoprecipitation - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Anti-PBR antibody [EPR5384] ab109497 (purified) at 1/60 immunoprecipitating PBR in U87-MG whole cell lysate.
Lane 1 (input): U87-MG whole cell lysate (10μg)
Lane 2 (+): Anti-PBR antibody [EPR5384] ab109497 + U87-MG whole cell lysate (10μg).
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PBR antibody [EPR5384] ab109497 in U87-MG whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (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 (Anti-PBR antibody [EPR5384] ab109497).
All lanes: Immunoprecipitation - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497)
Predicted band size: 19 kDa
Observed band size: 19 kDa
Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Anti-PBR antibody [EPR5384] 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 Anti-PBR antibody [EPR5384] ab109497 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Anti-PBR antibody [EPR5384] ab109497).
Immunocytochemistry/ Immunofluorescence - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling PBR with purified Anti-PBR antibody [EPR5384] ab109497 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 (Anti-PBR antibody [EPR5384] ab109497).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling PBR with unpurified Anti-PBR antibody [EPR5384] 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 (Anti-PBR antibody [EPR5384] ab109497).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-PBR antibody [EPR5384] - BSA and Azide free (ab213654)
Overlay histogram showing HepG2 cells stained with unpurified Anti-PBR antibody [EPR5384] 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 Anti-PBR antibody [EPR5384] ab109497, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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 (Anti-PBR antibody [EPR5384] ab109497).