Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all GPR18 Primary Antibodies »

Product Specifications

Immunogen

Partial synthetic peptide made to an N-terminal portion of the human GPR18 protein (between amino acids 1-50) [UniProt Q14330]

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for GPR18 Antibody - BSA Free

Western Blot: GPR18 AntibodyBSA Free [NBP2-24918]

Western Blot: GPR18 AntibodyBSA Free [NBP2-24918]

Western Blot: GPR18 Antibody [NBP2-24918] - Analysis of human spleen lysate (4 ug/mL) in the 1) absence and 2) presence of immunizing peptide 3) mouse spleen (6 ug/mL) and 4) rat spleen lysate (3 ug/mL) using GPR18 antibody.
Immunocytochemistry/ Immunofluorescence: GPR18 Antibody - BSA Free [NBP2-24918]

Immunocytochemistry/ Immunofluorescence: GPR18 Antibody - BSA Free [NBP2-24918]

Immunocytochemistry/Immunofluorescence: GPR18 Antibody [NBP2-24918] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-GPR18 Antibody NBP2-24918 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunohistochemistry-Paraffin: GPR18 Antibody - BSA Free [NBP2-24918]

Immunohistochemistry-Paraffin: GPR18 Antibody - BSA Free [NBP2-24918]

Immunohistochemistry-Paraffin: GPR18 Antibody [NBP2-24918] - Analysis of a FFPE tissue section of human tonsil using 1:200 dilution of GPR18 antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.
Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody [NBP2-24918] - An intracellular stain was performed on Daudi cells with GPR18 Antibody NBP2-24918 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Immunohistochemistry-Paraffin: GPR18 Antibody - BSA Free [NBP2-24918]

Immunohistochemistry-Paraffin: GPR18 Antibody - BSA Free [NBP2-24918]

Immunohistochemistry-Paraffin: GPR18 Antibody [NBP2-24918] - Analysis of human breast tumor tissue using an isotype control (top) and GPR18 antibody (bottom) at 5 ug/mL.
Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody [NBP2-24918] - An intracellular stain was performed on Ntera2 cells with GPR18 Antibody NBP2-24918 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody - BSA Free [NBP2-24918]

Flow Cytometry: GPR18 Antibody [NBP2-24918] - An intracellular stain was performed on THP-1 cells with GPR18 Antibody NBP2-24918 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Applications for GPR18 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 27256622)

Immunocytochemistry/ Immunofluorescence

2 - 10 ug/ml. Use reported in scientific literature (PMID 30894369)

Immunohistochemistry

1:200

Immunohistochemistry-Paraffin

1:200

Western Blot

2 ug/ml

Reviewed Applications

Read 1 review rated 3 using NBP2-24918 in the following applications:

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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

Store at -20C. Avoid freeze-thaw cycles.

Background: GPR18

GPR18 is an Orphan-A GPCR with no known ligand. GPR18 has been reported to be expressed at high levels in testis and spleen and at lower levels in other tissues associated with endocrine and immunologic/hematologic functions. ESTs have been isolated from human blood, bone marrow, brain, colon, liver/spleen, lymph, lymph node, testis, and tonsil libraries.

Long Name

G Protein-coupled Receptor 18

Alternate Names

GPCRW, NAGly Receptor

Gene Symbol

GPR18

Additional GPR18 Products

Product Documents for GPR18 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for GPR18 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.

Related Research Areas

Cancer Biomarkers

Citations for GPR18 Antibody - BSA Free

Customer Reviews for GPR18 Antibody - BSA Free (1)

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  • GPR18 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Jurkat and HT-29 human colon adenocarcinoma cell line
    Species: Human
    Verified Customer | Posted 01/23/2019
    Ladder - HT29 - Jurkat
    GPR18 Antibody - BSA Free NBP2-24918

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Protocols

View specific protocols for GPR18 Antibody - BSA Free (NBP2-24918):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

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FAQs for GPR18 Antibody - BSA Free

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  • Q: Has the antibody been tested in Flow Cytometry?

    A: Our GPR18 antibody with catalogue number NBP2-24918 has been tested for Western blotting and for IHC with paraffin-embedded tissue samples, but it has not yet been tested in any other applications. For your future reference, if we have tested one of our antibodies in a particular application and found it not to work, we would indicate this by writing (negative) after that application. In this instance you might wish to participate in our Innovator's Reward programme. This was created to allow researchers the opportunity to try our primary antibodies in an untested species or application, without the financial risk of failure. To participate you simply submit an online review detailing your positive or negative results. In return, you receive a discount voucher for 100% of the purchase price of the reviewed product. Further details can be found at the following link: Innovators Reward.

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