Overview

  • Product name
    Anti-G protein alpha Inhibitor 2 antibody
    See all G protein alpha Inhibitor 2 primary antibodies
  • Description
    Rabbit polyclonal to G protein alpha Inhibitor 2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Other Immunogen Type corresponding to Human G protein alpha Inhibitor 2.

Properties

Applications

Our Abpromise guarantee covers the use of ab20392 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
WB Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 40 kDa.
IP Use at an assay dependent dilution.
ICC/IF Use at an assay dependent dilution.

Target

  • Function
    Guanine nucleotide-binding proteins (G proteins) are involved as modulators or transducers in various transmembrane signaling systems. The G(i) proteins are involved in hormonal regulation of adenylate cyclase: they inhibit the cyclase in response to beta-adrenergic stimuli. May play a role in cell division.
  • Sequence similarities
    Belongs to the G-alpha family. G(i/o/t/z) subfamily.
  • Cellular localization
    Cytoplasm. Cytoplasm > cytoskeleton > centrosome. Cell membrane. Localizes in the centrosomes of interphase and mitotic cells. Detected at the cleavage furrow and/or the midbody.
  • Information by UniProt
  • Database links
  • Alternative names
    • Adenylate cyclase inhibiting G alpha protein antibody
    • Adenylate cyclase-inhibiting G alpha protein antibody
    • GIP antibody
    • GNAI 2 antibody
    • Gnai2 antibody
    • GNAI2_HUMAN antibody
    • GNAI2B antibody
    • GTP binding regulatory protein Gi alpha 2 chain antibody
    • Guanine nucleotide binding protein G protein alpha inhibiting activity polypeptide 2 antibody
    • Guanine nucleotide-binding protein G(i) subunit alpha-2 antibody
    • H LUCA15.1 antibody
    • H LUCA16.1 antibody
    • WUGSC:H LUCA15.1 antibody
    • WUGSC:H LUCA16.1 antibody
    see all

Images

  • Lanes 1 & 3 : Anti-G protein alpha Inhibitor 2 antibody (ab20392) at 1/1000 dilution
    Lane 2 : No primary

    All lanes : Rat RH7777 whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Lane 1 : No secondary
    Lanes 2-3 : Alkaline phosphatase conjugated goat anti-rabbit

    Performed under reducing conditions.

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 10 minutes

    See Abreview

References

This product has been referenced in:
  • Ramírez VT  et al. Wnt-5a/Frizzled9 Receptor Signaling through the Gao-Gß? Complex Regulates Dendritic Spine Formation. J Biol Chem 291:19092-107 (2016). Read more (PubMed: 27402827) »
  • Fu X  et al. MicroRNA-222-3p/GNAI2/AKT axis inhibits epithelial ovarian cancer cell growth and associates with good overall survival. Oncotarget 7:80633-80654 (2016). WB ; Human . Read more (PubMed: 27811362) »
See all 4 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question
Answer

Thank you for sending the image again.

As my colleague mentioned you may need to increase the denaturing and reducing conditions with this specific cell line.

My suggestion is to lysate the cells with RIPA buffer, a recipe for this buffer can be found from the following link:

https://docs.abcam.com/pdf/misc/abcam-protocols-book-2010.pdf

The lysis should be carried out on ice to reduce any degradation of the lysate.

Also, increasing the reducing conditions may also help. For that, heat the samples for longer in the presence of a reducing agent, such as DTT, which further denatures the proteins by reducing disulfide linkages, thus overcoming some forms of tertiary protein folding, and breaking up quaternary protein structure.

I would also suggest trying different blocking conditions, Have you tried blocking with 5% BSA for 1 or 2 hours at RT? If not, it could also be a potential optimisation step for this protocol.

Unfortunately, as we have not tried this particular cell line in our lab I am not able to provide definitive tips to get the expected results in your samples.

However, I hope this information has been of help. Do not hesitate to contact us for further assistance.

Read More

Answer

Thank you for your inquiry.

I am sorry to hear that you are getting results that are hard to interpret.

I believe the 120kDa band is not a complex containing G protein alpha Inhibitor 2 since you are using denaturing and reducing conditions. These conditions should break up all complexes.

If you think that the complex is not broken up I can suggest to increase the boiling time and also to try a different reducing agent.

Since the band is only appearing in one of the cell lysates, I believe that this must be a cell line specific artifact. Cell lines often have weird expression pattern if proteins and also acquire mutation over time. Since this band shows up at a very big size, it could be that the call line has a fusion protein with part of G protein alpha Inhibitor 2 protein. These kinds of fusion proteins are common in cancer cells lines and occur especially after prolonged culturing of the cell lines.

It would be of help to see an image of the results to better be able to make suggestions. Could you send one?

I also attached a Questionnaire to this email. This information will help me to see if I have any tips for optimizing the protocol. Could you please also include the cell line names and species?

I hope this information is helpful. please do not hesitate to contact me again with the requested detail if you have any further questions.

Read More

Question
Answer

Thank you for contacting us.

The information I was able to obtain regarding the immunogen is as follows. Any further details are considered proprietary.

The immunogen for this antibody is a synthetic peptide surrounding amino acid 229 of human Gai2. Since human Gai2 is a 355 aa protein,the immunogen is not located atthe extreme C-terminus.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (RH7777 Cells)
Loading amount
15 µg
Specification
RH7777 Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%

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Verified customer

Submitted Jul 31 2006

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