Overview

  • Product name
  • Description
    Rabbit polyclonal to RGS2
  • Host species
    Rabbit
  • Specificity
    PPlease note that a patterns of "background" bands may be present, this varies from tissue to tissue.
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    KREKMKRTLLKDWKTRC

    , corresponding to N terminal amino acids 29-44 of Human RGS2.

Properties

Applications

Our Abpromise guarantee covers the use of ab9963 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 1/10000 - 1/15000. Predicted molecular weight: 25 kDa.

Target

  • Function
    Regulates G protein-coupled receptor signaling cascades. Inhibits signal transduction by increasing the GTPase activity of G protein alpha subunits, thereby driving them into their inactive GDP-bound form (PubMed:11063746, PubMed:19478087). Plays a role in the regulation of blood pressure in response to signaling via G protein-coupled receptors and GNAQ. Plays a role in regulating the constriction and relaxation of vascular smooth muscle (By similarity). Binds EIF2B5 and blocks its activity, thereby inhibiting the translation of mRNA into protein (PubMed:19736320).
  • Tissue specificity
    Expressed in acute myelogenous leukemia (AML) and in acute lymphoblastic leukemia (ALL).
  • Sequence similarities
    Contains 1 RGS domain.
  • Post-translational
    modifications
    Phosphorylated by protein kinase C. Phosphorylation by PRKG1 leads to activation of RGS2 activity.
  • Cellular localization
    Cell membrane. Mitochondrion and Cell membrane. Cytoplasm. Nucleus, nucleolus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Basic helix-loop-helix phosphoprotein G0S8 antibody
    • Cell growth inhibiting protein 31 antibody
    • Cell growth-inhibiting gene 31 protein antibody
    • G0 to G1 switch regulatory 8 24kD antibody
    • G0/G1 switch regulatory protein 8 antibody
    • G0S8 antibody
    • GOS8 antibody
    • OTTHUMP00000060765 antibody
    • Regulator of G protein signaling 2 antibody
    • Regulator of G protein signalling 2 24kD antibody
    • Regulator of G-protein signaling 2 antibody
    • Regulators of G protein signaling XRGSVI antibody
    • RGS 2 antibody
    • RGS2 antibody
    • RGS2_HUMAN antibody
    see all

References

This product has been referenced in:
  • Snabaitis AK  et al. Regulation of the extracellular signal-regulated kinase pathway in adult myocardium: differential roles of G(q/11), Gi and G(12/13) proteins in signalling by alpha1-adrenergic, endothelin-1 and thrombin-sensitive protease-activated receptors. Cell Signal 17:655-64 (2005). WB ; Rat . Read more (PubMed: 15683740) »
  • Kveberg L  et al. Expression of regulator of G protein signalling proteins in natural killer cells, and their modulation by Ly49A and Ly49D. Immunology 115:358-65 (2005). Read more (PubMed: 15946253) »
See all 2 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Answer

Thank you for contacting us. I am sorry to hear that the antibody is giving you problems. As we discussed over the phone, potential troubleshooting tips that could help improve your results are: 1) loading more protein per lane (up to 100 ug max) 2) using more primary antibody (e.g. 1/1000) Please let me know if these tips were helpful or not. If they were not of help, I'd be happy to discuss a replacement, credit or refund with you. I wish you good luck and look forward to hear back from you!

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse ventricle)
Loading amount
50 µg
Specification
mouse ventricle
Gel Running Conditions
Reduced Denaturing (12% gradient gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Serban Georgescu

Verified customer

Submitted Apr 21 2008

Answer

I received confirmation today that the patterns of "background" bands varies from tissue to tissue. The VTA area of the brain has not been tested in the tester's hands therefore we do not have experience of a 12kDa band, but in other tissues other bands could be observed. I will update the datasheet and apologise I cannot provide more information regarding these bands, I hope this information will already help you,

Read More

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM I got the good size band plus a smaller one at about 14KDa. This aspecific band is stronger than the specific one. SAMPLE I used proteins extract from Ventral tegmenta area (Midbrain) PRIMARY ANTIBODY Antibody ab9963 diluted 1:10000 in 5%milk in PBS+0.2%Tween, incubation overnight at 4°C. 3*10 min wash in PBS+0.2%Tween DETECTION METHOD ECL Amersham POSITIVE AND NEGATIVE CONTROLS USED Check for actin for every samples ANTIBODY STORAGE CONDITIONS I aliquoted the antibody at the arrival and put the aliquots at -20. I used a new aliquot for every western blot SAMPLE PREPARATION The sample was put in a buffer conaining : 1M Tris pH8,150mM NaCl, 0.02%NaN3 10%, 0.1%SDS 10%, 1% NP40 ,0.5% Na deoxycholate, H2O, protease inhibitor cocktail "complete"(Sigma). The sample was mechanically brocken and quantified. Finally, i added SDS and the blue colorant to the sample, heated at 95°*10 min and loaded in the gel. AMOUNT OF PROTEIN LOADED 20 micrograms ELECTROPHORESIS/GEL CONDITIONS Gel used: NuPAGE 4-12% Bis-Tris Gel TRANSFER AND BLOCKING CONDITIONS Transfer : 1 h at 0.23 Amp in ice. Blocking : 1h30min in 5%milk in PBS+0.2%Tween SECONDARY ANTIBODY Antibody Goat ant rabbit HRP coniugated (Biorad), diluted 1:1000 in 5%milk in PBS+0.2%Tween. Incubation 45 min at room tempereture. 3*10 min wash in PBS+0.2%Tween HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 8 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No ADDITIONAL NOTES I found the second band in most of the case. I think this band is aspecific. To test it i would ask you if it's possible to replace this antibody with the other one you produce for RGS (ab14069). I am in the limits you ask for this remplacement. Thank you very much, best regards

Read More
Answer

Thank you for taking the time to submit these concerns to me. If there are only two bands on the blot and background this problem may be due in part to target breakdown or standard cross reactivity due to the fact that the product is a pAb. Since pAbs recognises multiple epitopes it could be the Ab pulling out an alternate target from your sample. You mention in the data forwarded that you have employed inhibitor cocktails. To prevent breakdown I would also advise that all sample preparation is carried out on ice, including mechanical homogenisation. If you are still concerned then you could locate a commercially available brain lysate which may suggest whether additional band is a peculiarity of brain sample. I would also like to advise that you run a no primary control in order to ascertain whether the band that you are seeing is due to the non specific binding of the secondary Ab rather than the primary.

Read More

Question

BATCH NUMBER 270602 ORDER NUMBER 05154 DESCRIPTION OF THE PROBLEM Using your suggestions the background was reduced but bands of a higher molecular weight were all that we could detect. SAMPLE Whole cell lysate of a 3T3 derived cell line shown to express the RNA PRIMARY ANTIBODY 1:2000 in PBST with no milk O/N at 4oC as suggested Previous blot: 1:2000 O/n 4oC with milk DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED We are running these blots blind as we lack a good positive (or negative ) control in the murine system. We are trying to get hold of human buffy coat PBMCs which are reported to express high levels of RGS2 but are still working on this. ANTIBODY STORAGE CONDITIONS overnight at 4oC. Following day aliquot and store in 5ul volumes at -20oC SAMPLE PREPARATION Sample prepared according to Marais et al., 1993. Proein estimation by Biorad reagent. Aliquot 30ug in 1X SB (120mM TRIS pH6.8; 20% glycerol;5% SDS and B-EtSH. Heat 5min at 95oC before loading. AMOUNT OF PROTEIN LOADED 30-50ug ELECTROPHORESIS/GEL CONDITIONS 12% PAGE/SDS gel TRANSFER AND BLOCKING CONDITIONS Electro blot Tris/Glycine/SDS with 20% MtOH (Dry Blot Buffer) for 2hrs at 20V. Blotting was successful as determined by the blot samples run in duplicate and the other half of the blot successfully probed with anti p27. Blocked as you suggested for 30min in PBST/5% milk. In the blot that had more background and putative band at the right size we blocked O/N in this mix. SECONDARY ANTIBODY Dako secondary anti rabbit - HRP diluted 1:3000 PBST no milk 1hr RT as suggested. Previous blot as above but with milk HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? Forst - I did the blot our way with the recommended concentration of RGS2 (1:10000) and got a completely negative blot. Then I increased the cell lysate to 50ug/track and tried 1:2000 dilution of RGS2 and got the blot with bands but very high background. I then contacted you and redid the blot exactly as you suggested (with 1:2000 RGS2)and on this occasion got less background, and multiple bands that were too high for RGS2. ADDITIONAL NOTES Will send scanned images to follow but our scanner isn't networked this morning.

Read More
Answer

Thank you for your patience. The originator of the antibody has never looked at 3T3 cells (which are fibroblasts), but at the RNA level RGS2 is most highly expressed in brain. In 3T3 cells, RGS2 mRNA may be low, or the protein may be unstable. The latter possibility is likely in view of Varshavsky's article, mentioned to me by the originator, in Science last year that described the regulation of RGS2 degradation. Although the originator of the antibody did not recall which RGS2 antibodies were used in this paper, the methods described therein as well as the paper itself could be excellent help for you. I believe you should use a brain lysate as a positive control. It is possible that the protein level in 3T3 cells is below detection.

Read More

Answer

Thank you for your phone call. I'm sending you a free of charge replacement for ab9963, for your record it is on order# 50879 and you should receive it Tuesday. The vial is from lot 27062 but I checked it and it is fine. Please let me know if you have any more questions and I do apologize for the inconvenience.

Read More

Question
Answer

Thank you for your enquiry. Ab9963 has been tested only against transfected or purified recombinant RGS2. If you have any more questions, please contact us again.

Read More

Answer

Thank you for your patience. Unfortunately I still have not heard from the originator of this antibody regarding your questions about the multiple bands. As for the background that you are experiencing, high background is usually a sign for blocking problem. You may want to try blocking with 3% BSA rather than milk. Also, the secondary may be reacting with the blocking reagent. I suggest that you test this by omitting the primary antibody and testing with only the secondary. If background appears, change either the secondary or the blocking reagent. Also, please try using the secondary at a lower concentration.

Read More

Answer

Thank you for your enquiry and your interest in our products. All the information available for this product is listed on the on-line datasheet (price, datasheet, publication, suitability, cross-reactivity). Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement.

Read More

Answer

Unfortunately, we do not have more information about this antibody. We will update the on-line datasheet of this product as soon as we get more data. We apologize for any inconvenience caused.

Read More

1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up