Anti-beta 1 Adrenergic Receptor antibody (ab103653)

Rabbit polyclonal beta 1 Adrenergic Receptor antibody. Validated in WB and tested in Human. Independently reviewed in 1 review(s). Immunogen corresponding to synthetic peptide.

Overview

  • Product name
    Anti-beta 1 Adrenergic Receptor antibody
    See all beta 1 Adrenergic Receptor primary antibodies
  • Description
    Rabbit polyclonal to beta 1 Adrenergic Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Guinea pig, Cat, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to a region within internal sequence amino acides 180-229 (CTVWAISALV SFLPILMHWW RAESDEARRC YNDPKCCDFV TNRAYAIASS) of Human beta 1 Adrenergic Receptor (NP_000675).

  • Positive control
    • Fetal Brain Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab103653 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 1 µg/ml. Predicted molecular weight: 51 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function
    Beta-adrenergic receptors mediate the catecholamine-induced activation of adenylate cyclase through the action of G proteins. This receptor binds epinephrine and norepinephrine with approximately equal affinity.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family. Adrenergic receptor subfamily. ADRB1 sub-subfamily.
  • Domain
    The PDZ domain-binding motif mediates competitive interactions with GOPC, MAGI3 and DLG4 and plays a role in subcellular location of the receptor.
  • Post-translational
    modifications
    Homologous desensitization of the receptor is mediated by its phosphorylation by beta-adrenergic receptor kinase.
  • Cellular localization
    Cell membrane. Localized at the plasma membrane. Found in the Golgi upon GOPC overexpression.
  • Information by UniProt
  • Database links
  • Alternative names
    • ADR B1 antibody
    • ADRB 1 antibody
    • ADRB1 antibody
    • ADRB1_HUMAN antibody
    • ADRB1R antibody
    • Adrenergic beta 1 receptor antibody
    • Adrenoceptor beta 1 antibody
    • B1AR antibody
    • Beta 1 adrenoceptor antibody
    • Beta 1 adrenoreceptor antibody
    • Beta-1 adrenergic receptor antibody
    • Beta-1 adrenoceptor antibody
    • Beta-1 adrenoreceptor antibody
    • BETA1AR antibody
    • RHR antibody
    see all

Images

  • Western blot analysis of Human fetal muscle tissue lysate labeling beta 1 Adrenergic Receptor with ab103653 at 1.0µg/ml.
  • Western blot analysis of Human fetal lung tissue lysate labeling beta 1 Adrenergic Receptor with ab103653 at 1.0µg/ml.
  • Western blot analysis of Human fetal liver tissue lysate labeling beta 1 Adrenergic Receptor with ab103653 at 1.0µg/ml.
  • Western blot analysis of Human fetal heart tissue lysate labeling beta 1 Adrenergic Receptor with ab103653 at 1.0µg/ml.
  • Western blot analysis of Human adult placenta tissue lysate labeling beta 1 Adrenergic Receptor with ab103653 at 1.0µg/ml.
  • Anti-beta 1 Adrenergic Receptor antibody (ab103653) at 1 µg/ml + Fetal Brain Lysate at 10 µg

    Predicted band size: 51 kDa



    Gel concentration: 12%

References

ab103653 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Q&A

Question

Dear, Our customer sent me answers and attached file.   -the human IgG administered, is it expected to increase the level of beta 1 Adrenergic Receptor in the cells tested? No, it is for our specific experiment. -I could not find reference to beta 1 Adrenergic Receptor being expressed in submandibular gland acinar cells, do you have any knowledge of its expression levels in this cell line? Do you have any access to a positive control (such as Fetal Brain Lysate) The cells used in our experiment are primary human submandibular acinar cells and these secretary cells have some canonical GPCRs expressed in the plasmamembrane such as M3 muscarinic receptor, α1-adrenergic receptor, β1-adrenergic receptor(Gordon B. Proctor, Guy H. Carpenter, Autonomic Neuroscience: Basic and Clinical 133 (2007) 3-18), which play an important role in salivary secretion. With the target protein (51.3 kDa) being so close in size to the IgG it may be a problem in resolving its presence. Especially with the protein potentially being post-translationally modified and with such high quantity of human IgG. You may have more success in using a secondary antibody which has been pre-adsorbed (such as ab97080 from our catalogue) or purifying your samples by performing IP prior to WB. Alternatively the human IgG could be removed by IP and the remaining sample loaded. We think there is a provable problem in the β1-AR antibody. We did some experiments and the results attached as a ppt file will be sent. In the results, you can see we even didn’t do IgG pretreatment (labeled as red Arabic numerals membrane), β1-AR antibody still can’t be detect as any considerable specific bands, however, other antibody (α1B-AR, from other company) was detected in the same protein loading condition. It’s interesting, even in our first experiment, in which we used human IgG as our specific experimental goal, α1B-AR antibody still was detected as specific bands, no IgG heavy chain bands was detected. It is true, because we used secondary antibodies for anti-goat in α1B-AR antibody case, and for anti-rabbit in β1-AR antibody case, why only in β1-AR antibody case anti-rabbit secondary antibody detected human IgG? By the way, this secondary antibodies was also used in other rabbit origin antibody western blot experiment (in which we also did human IgG pretreatment) and worked well.

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Answer

Thank you for providing all this information. You have clearly performed a set of carefully though out experiments and the ab103653 antibody does not appear to be producing the results expected. I thought the human IgG present in the samples may have been masking the signal but clearly this is not the case as you have performed the same experiment without the IgG present with no better result. It may be that you have unfortunately recieved a faulty vial of this antibody. In order to resolve this problem would you like to try an alternative lot of this antibody free of charge? Or an alternative antibody to the same target such as ab85037 or ab3442 which are both also rabbit polyclonals to beta 1 Adrenergic Receptor. Alternatively a credit note can be arranged. The reason why the anti-goat secondary antibody is not recognising the human IgG while the anti-rabbit antibody is probably down to the specificity of the antibodies used. If the antibody has been pre-adsorbed with serum from potentially cross reactive species this would reduce such background. You could try removing the human IgG prior to loading the gel through immunoprecipitation, or try using a different secondary antibody which has been preadsorbed. If you would like I can offer ab77189 which is a goat polyclonal to beta 1 Adrenergic Receptor which would allow you to use your anti-goat secondary antibody in your experiment. I look forward to hearing how you would like to proceed.

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Question

LOT NUMBER GR27166-3 ORDER NUMBER 946793 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE human submandibular gland acinar cells and we applied human IgG as treatments in the cells, which step we need in our experiment. PRIMARY ANTIBODY Concentration or dilution 1:1000 Diluent buffer TBST Incubation time overnight Incubation temperature: 4oC with shacking Wash Buffer TBST Number of washes 3 times, 10min/time, RT with shacking DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED - ANTIBODY STORAGE CONDITIONS -20C, We stored it according to the sheet. SAMPLE PREPARATION Lysis buffer: membrane fraction buffer: 100mM NaCl, 20mM Hepes, pH 7.4, 1mM MgCl2, 1mM dithiothreitol. Protease inhibitors: 0.3mM PMSF, 20µg/ml aprotinin, 1 µg/ml pepstatin A, 1 µg/ml leupetin Phosphatase inhibitors: did not use Reducing agent Boiling for ≥5 min? yes/no yes, 80oC 10min. AMOUNT OF PROTEIN LOADED 30ug/well ELECTROPHORESIS/GEL CONDITIONS 8% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Type of membrane PVDF Protein transfer verified Good Blocking agent and concentration 10% skim milk Blocking time 3h Blocking temperature RT SECONDARY ANTIBODY Species: goat Isotype: IgG Reacts against: rabbit Concentration or dilution 1:5000 Diluent buffer TBST Incubation time 1h Incubation temperature: RT Fluorochrome or enzyme conjugate: HRP Wash Buffer TBST Number of washes 3 times, 10min/time, RT with shacking HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Primary Ab dilution concentration (up to 1:750), blocking time(up to overnight), secondary diluent buffer, membrane washing buffer (3% skim milk in TBST), gel percentage (up to 12%). ADDITIONAL NOTES There was a too weak band for specific beta1-AR. I understand thick band at 55kDa, which is a nonspecific IgG heavy chain in my experiment. However, I don't understand what’s it at between 72kDa and 95kDa, which thicker even than specific band (at 51kDa, suggested by the sheet).

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Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would appreciate if you could confirm some further details: -the human IgG administered, is it expected to increase the level of beta 1 Adrenergic Receptor in the cells tested? -I could not find reference to beta 1 Adrenergic Receptor being expressed in submandibular gland acinar cells, do you have any knowledge of its expression levels in this cell line? Do you have any access to a positive control (such as Fetal Brain Lysate) With the target protein (51.3 kDa) being so close in size to the IgG it may be a problem in resolving its presence. Especially with the protein potentially being post-translationally modified and with such high quantity of human IgG. You may have more success in using a secondary antibody which has been pre-adsorbed (such as ab97080 from our catalogue) or purifying your samples by performing IP prior to WB. Alternatively the human IgG could be removed by IP and the remaining sample loaded. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I look forward to your reply

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