Overview

  • Product name
  • Description
    Goat polyclonal to GPCR GPR40
  • Host species
    Goat
  • Tested applications
    Suitable for: ELISA, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide:

    CAARTQGGKS QK

    , corresponding to C terminal amino acids 289-300 of Human GPCR GPR40 peptide.

  • Positive control
    • Human breast lysate

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris buffered saline, pH 7.3
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32889 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA 1/16000.
WB Use a concentration of 0.01 - 0.03 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 32 kDa).

Target

  • Function
    Receptor for medium and long chain saturated and unsaturated fatty acids. Binding of the ligand increase intracellular calcium concentration and amplify glucose-stimulated insulin secretion. The activity of this receptor is mediated by G-proteins that activate phospholipase C. Seems to act through a G(q) and G(i)-mediated pathway.
  • Tissue specificity
    Expressed abundantly in pancreatic beta cells.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • FFA1R antibody
    • FFAR 1 antibody
    • Ffar1 antibody
    • FFAR1_HUMAN antibody
    • Free fatty acid receptor 1 antibody
    • G protein coupled receptor 40 antibody
    • G-protein coupled receptor 40 antibody
    • GPCR40 antibody
    • GPR 40 antibody
    • GPR40 antibody
    see all

Images

  • Anti-GPCR GPR40 antibody (ab32889) at 0.03 µg/ml + Human breast lysate (35µg protein in RIPA buffer).

    Predicted band size: 32 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?



    Primary incubation was 1 hour.
    Detected by chemiluminescence.

References

ab32889 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Thank you for taking the time to contact us with your enquiry. I am very sorry you are having difficulty obtaining satisfactory results from this antibody (Ab32889) in western blotting. Below, I have copied a western blotting procedure used to test this antibody in the laboratory. I hope it will help. Bands of incorrect size are sometimes caused by inefficient denaturing and reducing of the protien. Your SDS (denaturing) sample buffer should contain a reducing agent (mercaptoethanol or DTT). This procedure mentions boiling for 5 minutes for denaturing and reducing, but you can try increasing this to 10 minutes if you have not already done so. Good luck with your experiments. If you have any more difficulty, please don't hesitate to get back to us. If you could please fill in the technical questionaire (see link below) and perhaps include and image of your blot, including a positive control and molecular weight markers, this would be very helpful. Thank you for your time. https://www.abcam.com/index.html?section=western&pageconfig=technical&mode=questionaire PROCEDURE: Tissue Lysis. Tissue chunks were weighed and cut into approx 1mm cubes using a razor blade. The tissue was transferred to a handheld homogenizer and 3 ml of ice-cold RIPA buffer was added per 1g of tissue. The tissue was gently homogenised over 20 minutes on ice. The resulting lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton X-100, 1% Sodium deoxycholate, 0.1% SDS). - SDS PAGE. Samples were run at 200V constant on a 12% acrylamide SDS-PAGE mini gel - using Biorad Mini-Protean 3 kit and protocols. Before loading samples had 5% (v/v) 2-ME added and were boiled for 3 minutes. - Transfer. We used a Biorad Mini Trans-Blot, constant 100 V for 1 hour. Transfer Buffer was 20 mM Tris pH 8.0, 150 mM Glycine, 10% Methanol. We transferred to PVDF membrane and stained with Ponceau Red to evaluate the transfer. - Staining. The membrane was blocked in 2.5% skimmed milk in TBS-T (TBS + 0.05% Tween-20) for 1 hr at room temperature with agitation. Primary antibody was incubated for 1 hr at room temperature with agitation. We used anti-goat-HRP secondary antibody for 1 hr at room temperature with agitation. We washed with TBST three times after primary and secondary antibody, each wash lasting for 5-10 mins. ECL-plus was used rather than ECL, which is considerably more sensitive. Final detection was on autoradiography film.

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