LPAR1/LPA1/EDG-2 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-03363

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Chicken

Cited:

Human, Mouse, Rat

Predicted:

Bovine (100%), Sheep (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Knockout Validated, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all LPAR1/LPA1/EDG-2 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to an internal portion of human EDG2 (within residues 200-300). [Swiss-Prot# Q92633]

Reactivity Notes

Predicted to react with sheep, and bovine based on 100% sequence homology. Positive staining in IHC-Fr/IF with 16 um cryostat sections of embryonic day 6 (E6) chicken brain was reported by a customer. Mouse reactivity reported in scientific literature (PMID: 24147068).

Localization

Cell membrane; Multi-pass membrane protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

43 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for LPAR1/LPA1/EDG-2 Antibody - BSA Free

Western Blot: LPAR1/LPA1/EDG-2 AntibodyBSA Free [NBP1-03363]

Western Blot: LPAR1/LPA1/EDG-2 AntibodyBSA Free [NBP1-03363]

Western Blot: LPAR1/LPA1/EDG-2 Antibody [NBP1-03363] - Detection of EDG2 in A549 whole cell extracts using NBP1-03363. The band at ~41 kDa position represents the target protein EDG2, whereas, the band at ~55kDa may potentially be the post-translationally modified (glycosylated, palmitoylated or lipidated) form of this protein.
Immunocytochemistry/ Immunofluorescence: LPAR1/LPA1/EDG-2 Antibody - BSA Free [NBP1-03363]

Immunocytochemistry/ Immunofluorescence: LPAR1/LPA1/EDG-2 Antibody - BSA Free [NBP1-03363]

Immunocytochemistry/Immunofluorescence: LPAR1/LPA1/EDG-2 Antibody [NBP1-03363] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-EDG2/LPAR1/LPA1 (NBP1-03363) at a 1:100 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with and anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunohistochemistry-Paraffin: LPAR1/LPA1/EDG-2 Antibody - BSA Free [NBP1-03363]

Immunohistochemistry-Paraffin: LPAR1/LPA1/EDG-2 Antibody - BSA Free [NBP1-03363]

Immunohistochemistry-Paraffin: LPAR1/LPA1/EDG-2 Antibody [NBP1-03363] - IHC analysis of formalin fixed paraffin-embedded (FFPE) human prostate cancer using LPAR1 antibody at 1:100 on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Staining was obseved in the scattered ducts. Staining was performed by Histowiz.
Simple Western: LPAR1/LPA1/EDG-2 AntibodyBSA Free [NBP1-03363]

Simple Western: LPAR1/LPA1/EDG-2 AntibodyBSA Free [NBP1-03363]

Simple Western: LPAR1/LPA1/EDG-2 Antibody [NBP1-03363] - Simple Western lane view shows a specific band for EDG2 in 0.5 mg/ml of A431 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for LPAR1/LPA1/EDG-2 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100-1:500

Immunohistochemistry

1:100-1:500

Immunohistochemistry-Frozen

1:100-1:500

Immunohistochemistry-Paraffin

1:100 -1:500

Simple Western

1:200

Western Blot

0.5 - 2.0 ug/ml
Application Notes
This EDG2 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry-Frozen and Western Blot, where a band is seen approx. 43 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in A431 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 43 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 1 review rated 4 using NBP1-03363 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: LPAR1/LPA1/EDG-2

Lysophosphatidic acid (LPA) is a natural small-molecule lipid of glycerophosphate originating from cellular membranes and it mainly acts through binding with its specific GPC-Receptors LPA1-5 (e.g. LPA receptor EDG-2 or LPA1/EDG2, EDG4 and EDG7) which can couple to Gi, Gq, G12, and possibly Gs for regulating specific downstream signal transduction pathways including those involving activation of purinergic receptors LPA6/P2Y5, GPR87 and P2Y10, TRPV1 and PPAR gamma. EDG2 interacts with RALA, ADRBK1 etc., and in the absence of LPA, it can be found predominantly at the cell surface whereas in the presence of LPA, it co-localizes with RALA in the endocytic vesicles. It is well established that platelets, certain inflammatory cells, nerve cells, endothelial cells, cancerous cells etc. are triggered to release LPA in autocrine/paracrine fashion with the induction of diverse biologic effects.

Long Name

Lysophosphatidic Acid Receptor 1

Alternate Names

EDG2, GPCR26, GPR26, LPA1, rec.1.3, vzg-1

Entrez Gene IDs

1902 (Human); 14745 (Mouse); 116744 (Rat)

Gene Symbol

LPAR1

UniProt

Q92633

Additional LPAR1/LPA1/EDG-2 Products

Product Documents for LPAR1/LPA1/EDG-2 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for LPAR1/LPA1/EDG-2 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 LPAR1/LPA1/EDG-2 Antibody - BSA Free

Customer Reviews for LPAR1/LPA1/EDG-2 Antibody - BSA Free (1)

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Customer Images

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  • Name: Eric Birgbauere
    Application: Immunofluorescence
    Sample Tested: 16 um cryostat sections of embryonic day 6 (E6) chicken brain
    Species: Other
    Verified Customer | Posted 10/20/2011
    LPAR1/LPA1/EDG-2 Antibody - BSA Free NBP1-03363

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Protocols

View specific protocols for LPAR1/LPA1/EDG-2 Antibody - BSA Free (NBP1-03363):

LPAR1/LPA1/EDG-2 Antibody:
Immunocytochemistry Protocol

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

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

LPAR1/LPA1/EDG-2 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-EDG-2 primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer 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.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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FAQs

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