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Cell Biology Apoptosis Receptors Associated Proteins

Anti-Adenosine A1 Receptor抗体(ab3460)

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Submit a review Q&A (4)References (4)

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Immunocytochemistry - Anti-Adenosine A1 Receptor antibody (ab3460)
  • Western blot - Anti-Adenosine A1 Receptor antibody (ab3460)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine A1 Receptor antibody (ab3460)

Key features and details

  • Rabbit polyclonal to Adenosine A1 Receptor
  • Suitable for: WB, ICC, IHC-P
  • Reacts with: Rat, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Adenosine A1 Receptor抗体
    参阅全部 Adenosine A1 Receptor 一抗
  • 描述

    兔多克隆抗体to Adenosine A1 Receptor
  • 宿主

    Rabbit
  • 特异性

    Detects Adenosine Receptor A1. This antibody does not detect other AR subtypes.
  • 经测试应用

    适用于: WB, ICC, IHC-Pmore details
  • 种属反应性

    与反应: Rat, Human
    预测可用于: Cow
  • 免疫原

    Synthetic peptide corresponding to Rat Adenosine A1 Receptor aa 309-326.
    Sequence:

    CQPKPPIDEDLPEEKAED


    (Peptide available as ab5893)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • WB: PC-12, SH-SY5Y, U-87MG, A549, K-562 whole cell lysates. ICC: HeLa cells IHC: rat brain tissue.
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.05% Sodium azide
    Constituents: 99% PBS, 0.1% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • Primary antibody说明

    Adenosine receptors (ARs) are members of the 7-transmembrane domain G-protein-coupled receptor superfamily. Structural, biochemical and pharmacological analyses of the AR genes and protein has led to the discovery of four distinct AR subtypes (A1, A2a, A2b, A3). Activation of ARs mediates several receptor subtype-specific physiological processes that include cardiac rate, smooth muscle tone, platelet aggregation, inflammation, cell growth and death, and neurotransmission. The A1AR is a glycoprotein that can activate Gi and Go proteins in vitro. In intact cells, agonist occupation of the A1AR has been shown to cause pertussis toxin-sensitive inhibition of adenylyl cyclase activity and, in some systems, a stimulation of phospholipase C resulting in mobilization of intracellular calcium stores. Activation of potassium channels by A1AR has been intensively studied in relation to its dramatic effects on the cardiovascular system. A1AR protein is highly expressed in brain (especially cerebellum, hippocampus, thalamus, and cortex) and spinal cord and in part, modulates neurotransmitter release. In white adipocytes A1AR inhibits lipolysis and stimulates glucose uptake. Other tissues also express A1AR including kidney and testis.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Apoptosis
    • Receptors
    • Associated Proteins
    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • GPCR
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Cardiovascular
    • Atherosclerosis
    • Ischemia / Reperfusion
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Cardiovascular
    • Heart
    • Cardiac arrhythmias
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Associated Proteins
    • Neuroscience
    • Processes

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Adenosine A1 Receptor peptide (ab5893)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3460于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/500. Predicted molecular weight: 37 kDa.Can be blocked with Adenosine A1 Receptor peptide (ab5893).
ICC
Use a concentration of 2 µg/ml.
IHC-P
1/50 - 1/200.
说明
WB
1/500. Predicted molecular weight: 37 kDa.Can be blocked with Adenosine A1 Receptor peptide (ab5893).
ICC
Use a concentration of 2 µg/ml.
IHC-P
1/50 - 1/200.

靶标

  • 功能

    Receptor for adenosine. The activity of this receptor is mediated by G proteins which inhibit adenylyl cyclase.
  • 序列相似性

    Belongs to the G-protein coupled receptor 1 family.
  • 细胞定位

    Cell membrane.
  • Target information above from: UniProt accession P30542 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 282133 Cow
    • Entrez Gene: 134 Human
    • Entrez Gene: 29290 Rat
    • Omim: 102775 Human
    • SwissProt: P28190 Cow
    • SwissProt: P30542 Human
    • SwissProt: P25099 Rat
    • Unigene: 77867 Human
    • Unigene: 32078 Rat
    see all
  • 别名

    • A1AR antibody
    • A1R antibody
    • AA1R antibody
    • AA1R_HUMAN antibody
    • Adenosine receptor A1 antibody
    • ADORA 1 antibody
    • ADORA 1 antibody
    • ADORA1 antibody
    • RDC 7 antibody
    • RDC7 antibody
    • Ri antibody
    see all

图片

  • Immunocytochemistry - Anti-Adenosine A1 Receptor antibody (ab3460)
    Immunocytochemistry - Anti-Adenosine A1 Receptor antibody (ab3460)

    Immunocytochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton™ X-100 permeabilized HeLa cells staining adenosine A1 receptor with ab3460 at 2 µg/ml and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution (green). Nuclei are stained with SlowFade® Gold Antifade Mountant with DAPI  (blue) and F-actin is stained with Alexa Fluor® 555 Rhodamine Phalloidin (red). Negative control contains no primary anibody.

  • Western blot - Anti-Adenosine A1 Receptor antibody (ab3460)
    Western blot - Anti-Adenosine A1 Receptor antibody (ab3460)
    All lanes : Anti-Adenosine A1 Receptor antibody (ab3460) at 1/250 dilution

    Lane 1 : PC-12 whole cell extract
    Lane 2 : SH-SY5Y whole cell extract
    Lane 3 : U-87 MG whole cell extract
    Lane 4 : A549 whole cell extract
    Lane 5 : K-562 whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 37 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine A1 Receptor antibody (ab3460)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenosine A1 Receptor antibody (ab3460)

    ab3460 labelling Adenosine A1 receptor in the cytoplasm of Rat brain tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

实验方案

  • Immunohistochemistry protocols

Click here to view the general protocols

数据表及文件

  • Datasheet download

    Download

文献 (4)

发表研究结果有使用 ab3460?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab3460 被引用在 4 文献中.

  • Rivera-Oliver M  et al. Adenosine A1-Dopamine D1 Receptor Heteromers Control the Excitability of the Spinal Motoneuron. Mol Neurobiol 56:797-811 (2019). PubMed: 29797183
  • Stettler O  et al. Engrailed homeoprotein recruits the adenosine A1 receptor to potentiate ephrin A5 function in retinal growth cones. Development 139:215-24 (2012). PubMed: 22147955
  • Datino T  et al. Mechanisms by which adenosine restores conduction in dormant canine pulmonary veins. Circulation 121:963-72 (2010). WB ; Dog . PubMed: 20159830
  • Kempson SA  et al. Acute inhibition of the betaine transporter by ATP and adenosine in renal MDCK cells. Am J Physiol Renal Physiol 295:F108-17 (2008). WB ; Dog . PubMed: 18448594

客户评价及客户问答

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1-4 of 4 Abreviews or Q&A

Question

I am using at the moment your anti-adenosine A1 receptor antibody (ab3460) for WB on dog heart tissue. I have got a lot of unspecific bands and I did not get a nice band at 37 kDa. Do you have any positive control of dog A1 adenosine receptor? What protocol for protein isolation and WB would you suggest to improve my results?

Read More

Abcam community

Verified customer

Asked on Jun 15 2012

Answer

Following up my previous message, the immunogen for ab124780, an alternative to ab3460, is also unfortunately unlikely to react with canine Adenosine A1 Receptor. Alignment of the canine receptor amino acid sequence with that of the ab124780 immunogen returns 60% identity. We generally consider this to indicate a low probability of cross-reaction.

Please let me know if we can help in any way.

Read More

Abcam Scientific Support

回复于 Jun 15 2012

Question

I am using at the moment your anti-adenosine A1 receptor antibody (ab3460) for WB on dog heart tissue. I have got a lot of unspecific bands and I did not get a nice band at 37 kDa. Do you have any positive control of dog A1 adenosine receptor? What protocol for protein isolation and WB would you suggest to improve my results?

Read More

Abcam community

Verified customer

Asked on Jun 14 2012

Answer

Thank you for contacting us. I think the issue is probably that the homology between the immunogen and the canine receptor is low: 67% identity. We do not have a dog AAR1 to test this.

In general, for multi-pass membrane receptors, we suggest not boiling the sample before loading into the gel , to discourage the aggregation that boiling can induce. Heating at 70C for 10 minutes should be sufficient to denature the sample. (Aggregates do not enter the gel easily). Otherwise, the protocol should be standard, blocking with 5% milk or BSA, and incubation with the antibody diluted to 1/500 in TBST and incubating overnight at 4C. We suggest loading 20-50ug of sample per lane.

Have you considered any of the other AAR1 antibodies we have? None have been tested for reactivity with dog but I will check the immunogen for ab124780 against the dog sequence (http://www.uniprot.org/uniprot/P11616.fasta) and will send the result of the alignment when I have it.

Read More

Abcam Scientific Support

回复于 Jun 14 2012

Question

Many thanks for your reply to my query. I have tried this antibody in brain and found that the neurones are adequately demonstrated as is heart muscle. However, the frozen section results all appear somewhat stronger than the paraffin wax samples. My complaint was that RTPCR data suggests that A1 receptors are present in the smooth muscle around airways. This appeared to be the case with frozen lung sections but no staining was achieved when paraffin wax sections were employed. We used a standard microwaving protocol using pH6 citrate buffer. Have you any other suggestions or do you feel that the expression levels are much lower in lung compared with brain? I look forward to your comments. Regards,

Read More

Abcam community

Verified customer

Asked on Aug 02 2004

Answer

Thank you for your reply. If the signal is stronger on frozen section compared to formalin-fixed sections, it indicates that the epitope unmasking may not be proper. We would suggest trying to treat the samples in microwave for different times and to see if the intensity of the signal improves. It is know that certain tissue requires longer retrieval than the other and even if you use a standard well characterized protocol, it may be worth optimizing the technique. As you have suggested, it may well be that the expression level is lower in the lung. One comment on the RT-PCR results - you may need to handle RT-PCR data very carefully. My experience is that high RNA levels do not always correspond with high levels of transcribed protein. If you need anything further or any help then please let us know.

Read More

Abcam Scientific Support

回复于 Aug 04 2004

Question

ANTIBODY CODE 3460 DESCRIPTION OF THE PROBLEM No obvious specific staining in paraffin sections, just general cell staining with no obvious smooth muscle staining except in vasculature smooth muscle staining in cryostat sections together with some nuclear chromatin staining SAMPLE Human lung PRIMARY ANTIBODY ABCAM/ Human / PBS / 1/100;1/400; 1/800; / 1hour / PBS SECONDARY ANTIBODY Goat anti rabbit biotinylated DETECTION METHOD DAB POSITIVE AND NEGATIVE CONTROLS USED IgG controls FIXATION OF SAMPLE Paraformaldehyde fixed overnight, then paraffin processed Frozen sections fixed in paraformaldehyde for 2 minutes ANTIGEN RETRIEVAL Paraffin sections microwaved in citrate buffer for 20 minutes at 100C, then allowed to cool for 20 minutes No retrieval used for frozens PERMEABILIZATION STEP No permeabilization 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? dilution ADDITIONAL NOTES In lung the A1 receptor is known to be expressed in smooth muscle. we could demonstrate this in frozen sections but not in paraffin processed sections which is what the antibody claimed to be able to do. What pretreatments do you use to get it to work in paraffins and what is causing the chromatin staining in the frozen sections

Read More

Abcam community

Verified customer

Asked on Jul 22 2004

Answer

Thank you for your enquiry and for your patience. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. A1AR protein is highly expressed in brain (especially cerebellum, hippocampus, thalamus, and cortex) and spinal cord and in part, modulates neurotransmitter release. This antibody was tested in human brain (hippocampus sections) and the pattern is plasma membrane staining. Therefore we would suggest using positive control (i.e. human or rat brain sections) along with the samples. If you use formalin-fixed sections we would suggest antigen retrieval (microwave method), but please make sure that the samples are not over-treated. We would also suggest blocking endogenous peroxidase activity which may give unspecific staining. If you are still having problem with this antibody, please contact us again providing the batch number.

Read More

Abcam Scientific Support

回复于 Aug 02 2004

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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