GPR81 Antibody - BSA Free
Novus Biologicals | Catalog # NLS2095
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Key Product Details
Species Reactivity
Validated:
Human, Mouse
Cited:
Human, Mouse, Rat
Predicted:
Primate (100%). Backed by our 100% Guarantee.
Applications
Validated:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence
Cited:
Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
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Product Specifications
Immunogen
Synthetic 19 amino acid peptide from C-terminus of human GPR81.
Epitope
C-Terminus
Reactivity Notes
Predicted cross-reactivity based on sequence identity: Gorilla (100%), Marmoset (95%). Mouse reactivity reported in the literature (PMID: 19633298).
Localization
Multi-pass membrane protein
Specificity
Human GPR81. BLAST analysis of the peptide immunogen showed no homology with other human proteins, except KRBA1 (53%).
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Description
Product can be stored undiluted at 4C for up to 1 month.
Scientific Data Images for GPR81 Antibody - BSA Free
Immunocytochemistry/ Immunofluorescence: GPR81 Antibody - BSA Free [NLS2095]
GPR81-Antibody-Immunocytochemistry-Immunofluorescence-NLS2095-img0010.jpgImmunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Brain, Substantia Nigra, pigmented and nonpigmented neurons.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Analysis of anti-FKSG80 / GPR81 antibody with human anterior pituitary.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Analysis of anti-FKSG80 / GPR81 antibody with human colon, carcinoma.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Human Skin, Melanoma formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Human Ovary, Carcinoma formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Human pancreas, carcinoma formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.Immunohistochemistry-Paraffin: GPR81 Antibody - BSA Free [NLS2095]
Immunohistochemistry-Paraffin: GPR81 Antibody [NLS2095] - Breast, CarcinomaWestern Blot: GPR81 Antibody - BSA Free [NLS2095] -
Western Blot: GPR81 Antibody - BSA Free [NLS2095] - Stable knockdown of GPR81 in MDA-MB-231 cells. Using a lentiviral system, GPR81 was stably knocked down in MDA-MB-231 cells with GPR81-specific short hairpin RNA (shGPR81 #1) & compared with cells containing scrambled shRNA as a control (shNT). Knockdown of GPR81 was confirmed by (a) western blotting & (b) RT-qPCR. The GPR81, MCT1, & MCT4 mRNA levels in the shGPR81 #1 cells are presented as the fold increase compared with those in shNT cells. Data are presented as the means ± s.d. (n = 3/group). **p < 0.01; Student’s t-test. (c) Lactate production in shNT & shGPR81 #1 cells. The cells were cultured for 24 h & lactate concentrations in the supernatants were determined (n = 3/group). **p < 0.01 (vs. shNT cells); Student’s t-test. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35428832), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: GPR81 Antibody - BSA Free [NLS2095] -
Western Blot: GPR81 Antibody - BSA Free [NLS2095] - Expression of GPR81 in breast cancer cells. (a) Western blot analysis of GPR81 in breast cancer cells (MCF-7 & MDA-MB-231) & non-tumorigenic epithelial cells (MCF-10A). Cell lysates were analyzed by immunoblotting using anti-GPR81, anti-MCT4, anti-MCT1, & anti-beta -actin antibodies. (b) Immunocytochemical analysis of GPR81 expression in breast cancer cells (MCF-7 & MDA-MB-231) & non-tumorigenic epithelial cells (MCF-10A). MCF-10A, MCF-7 & MDA-MB-231 cells were incubated with polyclonal antibodies against GPR81 followed by Alexa Fluor Plus 488-conjugated secondary antibody & visualized under a fluorescence microscope. Na/K ATPase was used as a cell membrane marker & the nuclei were stained with DAPI. Scale bar 10 μm. (c) MDA-MB-231 cells were cultured in the presence & absence of 20 mM lactate for 48 h, & cell lysates were analyzed by immunoblotting using anti-GPR81 & anti-beta -actin antibodies. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35428832), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Western Blot: GPR81 Antibody - BSA Free [NLS2095] -
Western Blot: GPR81 Antibody - BSA Free [NLS2095] - Expression of GPR81 in breast cancer cells. (a) Western blot analysis of GPR81 in breast cancer cells (MCF-7 & MDA-MB-231) & non-tumorigenic epithelial cells (MCF-10A). Cell lysates were analyzed by immunoblotting using anti-GPR81, anti-MCT4, anti-MCT1, & anti-beta -actin antibodies. (b) Immunocytochemical analysis of GPR81 expression in breast cancer cells (MCF-7 & MDA-MB-231) & non-tumorigenic epithelial cells (MCF-10A). MCF-10A, MCF-7 & MDA-MB-231 cells were incubated with polyclonal antibodies against GPR81 followed by Alexa Fluor Plus 488-conjugated secondary antibody & visualized under a fluorescence microscope. Na/K ATPase was used as a cell membrane marker & the nuclei were stained with DAPI. Scale bar 10 μm. (c) MDA-MB-231 cells were cultured in the presence & absence of 20 mM lactate for 48 h, & cell lysates were analyzed by immunoblotting using anti-GPR81 & anti-beta -actin antibodies. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35428832), licensed under a CC-BY license. Not internally tested by Novus Biologicals.Applications for GPR81 Antibody - BSA Free
Application
Recommended Usage
Immunohistochemistry-Paraffin
2.5 - 20 ug/ml
Application Notes
WB reactivity reported in (PMID: 19633298). ICC/IF reactivity reported in (PMID: 32071396)..
Formulation, Preparation, and Storage
Purification
Immunogen affinity purified
Formulation
PBS
Format
BSA Free
Preservative
0.1% Sodium Azide
Concentration
1.0 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Keep as concentrated solution. Aliquot and store at -20C or below. Avoid multiple freeze-thaw cycles.
Background: GPR81
Long Name
G Protein-coupled Receptor 81
Alternate Names
FKSG80, GPR104, HCA1, HCAR1, LACR1, TA-GPCR
Entrez Gene IDs
27198 (Human)
Gene Symbol
HCAR1
UniProt
Additional GPR81 Products
Product Documents for GPR81 Antibody - BSA Free
Product Specific Notices for GPR81 Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Citations for GPR81 Antibody - BSA Free
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Protocols
View specific protocols for GPR81 Antibody - BSA Free (NLS2095):
Immunohistochemistry Protocol for GPR81 Antibody (NLS2095):
Immunohistochemistry
1. Prepare tissue with formalin fixation and by embedding it in paraffin wax.
2. Make 4 um sections and place on pre-cleaned and charged microscope slides.
3. Heat in a tissue-drying oven for 45 minutes @ 60 degrees Celcius.
4. Deparaffinize the tissues by wash drying the slides in 3 changes of xylene for 5 minutes each @ RT.
5. Rehydrate the tissues by washing the slides in 3 changes of 100% alcohol for 3 minutes each @ RT.
6. Wash the slides in 2 changes of 95% alcohol for 3 minutes each @ RT.
7. Wash the slides in 1 change of 80% alcohol for 3 minutes @ RT.
8. Rinse the slides in gentle running distilled water for 5 minutes @ RT.
9. Perform antigen retrieval by steaming the slides in 0.01M sodium citrate buffer (pH 6.0) @ 99-100 degrees Celcius
for 20 minutes.
10. Remove the slides from the heat and let stand in buffer @ RT for 20 minutes.
11. Rinse the slides in 1X TBS-T for 1 minute @ RT.
**Do not allow the tissues to dry at any time during the staining procedure**
12. Begin the immunostaining by applying a universal protein block for 20 minutes @ RT.
13. Drain protein block from the slides and apply the diluted primary antibody for 45 minutes @ RT.
14. Rinse the slide in 1X TBS-T for 1 minute @ RT.
15. Apply a biotinylated anti-rabbit IgG (H+L) secondary for 30 minutes @ RT.
16. Rinse the slide in 1X TBS-T for 1 minute @ RT.
17. Apply an alkaline phosphatase steptavidin for 30 minutes @ RT.
18. Rinse the slide in 1X TBS-T for 1 minute @ RT.
19. Apply an alkaline phosphatase chromagen substrate for 30 minutes @ RT.
20. Rinse the slide in distilled water for 1 minute @ RT.
**This method should only be used if the chromagen substrate is alcohol insoluble (ie: Vector Red, DAB)**
21. Dehydrate the tissue by washing the slides in 2 changes of 80% alcohol for 1 minute each @ RT.
22. Wash the slides in 2 changes of 95% alcohol for 1 minute each @ RT.
23. Wash the slides in 3 changes of 100% alcohol for 1 minute each @ RT.
24. Wash the slides in 3 changes of xylene for 1 minute each @ RT.
25. Apply cover slip.
Immunohistochemistry
1. Prepare tissue with formalin fixation and by embedding it in paraffin wax.
2. Make 4 um sections and place on pre-cleaned and charged microscope slides.
3. Heat in a tissue-drying oven for 45 minutes @ 60 degrees Celcius.
4. Deparaffinize the tissues by wash drying the slides in 3 changes of xylene for 5 minutes each @ RT.
5. Rehydrate the tissues by washing the slides in 3 changes of 100% alcohol for 3 minutes each @ RT.
6. Wash the slides in 2 changes of 95% alcohol for 3 minutes each @ RT.
7. Wash the slides in 1 change of 80% alcohol for 3 minutes @ RT.
8. Rinse the slides in gentle running distilled water for 5 minutes @ RT.
9. Perform antigen retrieval by steaming the slides in 0.01M sodium citrate buffer (pH 6.0) @ 99-100 degrees Celcius
for 20 minutes.
10. Remove the slides from the heat and let stand in buffer @ RT for 20 minutes.
11. Rinse the slides in 1X TBS-T for 1 minute @ RT.
**Do not allow the tissues to dry at any time during the staining procedure**
12. Begin the immunostaining by applying a universal protein block for 20 minutes @ RT.
13. Drain protein block from the slides and apply the diluted primary antibody for 45 minutes @ RT.
14. Rinse the slide in 1X TBS-T for 1 minute @ RT.
15. Apply a biotinylated anti-rabbit IgG (H+L) secondary for 30 minutes @ RT.
16. Rinse the slide in 1X TBS-T for 1 minute @ RT.
17. Apply an alkaline phosphatase steptavidin for 30 minutes @ RT.
18. Rinse the slide in 1X TBS-T for 1 minute @ RT.
19. Apply an alkaline phosphatase chromagen substrate for 30 minutes @ RT.
20. Rinse the slide in distilled water for 1 minute @ RT.
**This method should only be used if the chromagen substrate is alcohol insoluble (ie: Vector Red, DAB)**
21. Dehydrate the tissue by washing the slides in 2 changes of 80% alcohol for 1 minute each @ RT.
22. Wash the slides in 2 changes of 95% alcohol for 1 minute each @ RT.
23. Wash the slides in 3 changes of 100% alcohol for 1 minute each @ RT.
24. Wash the slides in 3 changes of xylene for 1 minute each @ RT.
25. Apply cover slip.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for GPR81 Antibody - BSA Free
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A: This antibody was raised to a synthetic 19 amino acid peptide derived from the C-terminus of the human protein. Human GPR81 has a theoretical molecular weight of approximately 39kDa according to the Uniprot accession: Uniprot site
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