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Neuroscience Neurotransmission Receptors / Channels GPCR Cannabinoid Receptor

Anti-Cannabinoid Receptor I抗体(ab3558)

  • Datasheet
  • SDS
Reviews (1)Q&A (14)References (3)

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Immunocytochemistry/ Immunofluorescence - Anti-Cannabinoid Receptor I antibody (ab3558)

    Key features and details

    • Rabbit polyclonal to Cannabinoid Receptor I
    • Suitable for: IP, ICC/IF, IHC-Fr, IHC-P, Flow Cyt, WB
    • Reacts with: Mouse, Rat, Rabbit, Human, Ferret
    • Isotype: IgG

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    概述

    • 产品名称

      Anti-Cannabinoid Receptor I抗体
      参阅全部 Cannabinoid Receptor I 一抗
    • 描述

      兔多克隆抗体to Cannabinoid Receptor I
    • 宿主

      Rabbit
    • 特异性

      Detects CB1 from Human and Rat tissues as well as transfected Rat CB1.
    • 经测试应用

      适用于: IP, ICC/IF, IHC-Fr, IHC-P, Flow Cyt, WBmore details
    • 种属反应性

      与反应: Mouse, Rat, Rabbit, Human, Ferret
      预测可用于: Cat, Chimpanzee, Rhesus monkey
    • 免疫原

      Recombinant fragment within Human Cannabinoid Receptor I aa 1-100 (N terminal). The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.

    • 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 or -80°C. Avoid freeze / thaw cycle.
    • 存储溶液

      Preservative: 0.05% Sodium azide
      Constituents: 50% Glycerol (glycerin, glycerine), 0.1% BSA, 49% PBS
    • Concentration information loading...
    • 纯度

      Immunogen affinity purified
    • 克隆

      多克隆
    • 同种型

      IgG
    • 研究领域

      • Neuroscience
      • Neurotransmission
      • Receptors / Channels
      • GPCR
      • Cannabinoid Receptor
      • Cardiovascular
      • Lipids / Lipoproteins
      • Lipid Metabolism
      • Cholesterol Metabolism
      • Cardiovascular
      • Atherosclerosis
      • Lipoprotein metabolism
      • Metabolism
      • Pathways and Processes
      • Metabolic signaling pathways
      • Lipid and lipoprotein metabolism
      • Cholesterol Metabolism
      • Metabolism
      • Pathways and Processes
      • Metabolic signaling pathways
      • Lipid and lipoprotein metabolism
      • Lipoprotein metabolism
      • Metabolism
      • Types of disease
      • Cancer
      • Metabolism
      • Types of disease
      • Heart disease
      • 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)
    • Isotype control

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

      • Recombinant Human Cannabinoid Receptor I protein (ab158150)

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab3558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
    IP
    Use at an assay dependent concentration. PubMed: 22387618
    ICC/IF
    1/1000.
    IHC-Fr
    Use at an assay dependent concentration.
    IHC-P
    Use at an assay dependent concentration.
    Flow Cyt
    Use at an assay dependent concentration. PubMed: 20206138

    ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

    WB
    Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.

    By Western blot, this antibody detects an ~60 kDa protein representing CB1 from rat brain homogenate.

    说明
    IP
    Use at an assay dependent concentration. PubMed: 22387618
    ICC/IF
    1/1000.
    IHC-Fr
    Use at an assay dependent concentration.
    IHC-P
    Use at an assay dependent concentration.
    Flow Cyt
    Use at an assay dependent concentration. PubMed: 20206138

    ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

    WB
    Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.

    By Western blot, this antibody detects an ~60 kDa protein representing CB1 from rat brain homogenate.

    靶标

    • 功能

      Involved in cannabinoid-induced CNS effects. Acts by inhibiting adenylate cyclase. Could be a receptor for anandamide. Inhibits L-type Ca(2+) channel current. Isoform 2 and isoform 3 have altered ligand binding.
    • 组织特异性

      Widely expressed.
    • 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 P21554 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: 1268 Human
      • Entrez Gene: 12801 Mouse
      • Entrez Gene: 25248 Rat
      • Omim: 114610 Human
      • SwissProt: O02777 Cat
      • SwissProt: Q5IS73 Chimpanzee
      • SwissProt: P21554 Human
      • SwissProt: P47746 Mouse
      • SwissProt: P20272 Rat
      • SwissProt: Q71SP5 Rhesus monkey
      • Unigene: 709067 Human
      • Unigene: 75110 Human
      • Unigene: 7992 Mouse
      • Unigene: 222240 Rat
      • Unigene: 89774 Rat
      see all
    • 别名

      • CANN6 antibody
      • Cannabinoid receptor 1 antibody
      • CB-R antibody
      • CB1 antibody
      • CB1A antibody
      • CB1K5 antibody
      • CB1R antibody
      • Central cannabinoid receptor antibody
      • CNR antibody
      • CNR1 antibody
      • CNR1_HUMAN antibody
      • OTTHUMP00000016838 antibody
      • OTTHUMP00000214579 antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-Cannabinoid Receptor I antibody (ab3558)
      Immunocytochemistry/ Immunofluorescence - Anti-Cannabinoid Receptor I antibody (ab3558)

      ab3558 at a 1:1000 dilution staining Rat CB1 in transfected AtT20 cells by immunocytochemistry.

    实验方案

    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (3)

    发表研究结果有使用 ab3558?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3558 被引用在 3 文献中.

    • Kleyer J  et al. Cannabinoid receptor trafficking in peripheral cells is dynamically regulated by a binary biochemical switch. Biochem Pharmacol 83:1393-412 (2012). Flow Cyt, IP ; Human . PubMed: 22387618
    • Shen L  et al. The role of peripheral cannabinoid receptors type 1 in rats with visceral hypersensitivity induced by chronic restraint stress. J Neurogastroenterol Motil 16:281-90 (2010). WB ; Rat . PubMed: 20680167
    • Leonti M  et al. Falcarinol is a covalent cannabinoid CB1 receptor antagonist and induces pro-allergic effects in skin. Biochem Pharmacol 79:1815-26 (2010). WB, Flow Cyt ; Human . PubMed: 20206138

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 15 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cannabinoid Receptor I antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (adult brain)
    Specification
    adult brain
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Carl Hobbs

    Verified customer

    提交于 Oct 16 2007

    Question

    if we had a CB1 and CB2 antibody which was appropriate for Image flow cytometry. She was interested in purchasing both a primary and secondary antibody. She is using PMBC cells and has done the gene expression for the proteins she is looking at.

    Thank you

    Read More

    Abcam community

    Verified customer

    Asked on Jul 31 2012

    Answer

    Thank you for contacting us.

    We have following CB1 and CB2 available. These are testes in flow cytometry and are fully guaranteed.

    ab3558; https://www.abcam.com/Cannabinoid-Receptor-I-antibody-ab3558.html

    ab3561; https://www.abcam.com/Cannabinoid-Receptor-II-antibody-ab3561.html

    ab3560; https://www.abcam.com/Cannabinoid-Receptor-II-antibody-ab3560.html

    Could you let me know which conjugated you would like for compatible secondary antibody?

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

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Jul 31 2012

    Question

    We are using ab3558 and ab3560 for flow cytometry and would like to know which isotype control you recommend.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 21 2012

    Answer

    Thank you for your inquiry.

    The best way to find isotype controls for an antibody is to start at our homepage www.abcam.com. Go to the yellow bar at the top of the page and move the mouse to “Products,” then click on “Isotype Controls”. The direct link is:

    https://www.abcam.com/index.html?pageconfig=catalog_byproducttype&intProductTypeID=20

    Then select the correct isotype control based on the host species as well as the isotype, sub-isotype, conjugation, and application of the primary antibody which you are using.

    For example, if the primary antibody is a FITC-conjugated mouse IgG1 for use in flow cytometry, then you will need to choose a FITC-conjugated mouse IgG1 isotype control tested in flow cytometry.

    Please note that most isotype controls are monoclonal. They are not suitable for use with polyclonal antibodies as these contain more than one IgG subclass. However, we do provide several polyclonal isotype controls for use in these circumstances.

    I taken the information that you have provided me to determine that the best isotype control in this instance is ab37415 Rabbit IgG- Chip Grade. Should you decide to conjugate a fluorescent tag onto your antibodies however,it is advised that you use a different isotype control which is also conjugated to the same fluorescent label.

    Please let me know if have any questions. I would be happy to assist you.

    Read More

    Abcam Scientific Support

    回复于 Feb 21 2012

    Question

    Please provide a western blot image for ab3559 and ab3558

    Read More

    Abcam community

    Verified customer

    Asked on Apr 22 2008

    Answer

    Thank you for your enqiury. According to our most recent internal testing data, by Western blot, ab3558 Cannabinoid Receptor I antibody shows detection of an ~ 52 kDa band in rat brain homogenate. After further research on anti-Cannabinoid Receptor 1 ab3558 , some recently published articles have shed light on the molecular weight. Reference 1 shows this antibody detecting CB1 at ~50 kDa (see reference 1 below). However, a more recent reference shows our antibody detecting CB1 expressed in macaque cortical tissue homogenate at a molecular weight of ~85 kDa and ~52 kDa (see reference 2 below). Therefore, according to the literature, it appears that ab3558 Cannabinoid Receptor I antibody will recognize CB1 at two different molecular weights, ~50 kDa and ~85 kDa. References: Reference 1: Citation: J. Neurosci., December 3, 2003. 23(35):11136-11141 Link: http://www.jneurosci.org/cgi/reprint/23/35/11136 Description: This article cites ABR's anti-Cannabinoid Receptor 1 (PA1-743) being used in WB on temporal cortex from human AD patients. "Western blotting. The protocol used is basically as described previously (Romero et al., 2002). Human brain was obtained at autopsy and a 1 gm piece of cerebral cortical gray matter was homogenized in 10 ml of M-PER mammalian protein extraction reagent. The homogenate was shaken gently for 10 min and then centrifuged at 27,000 x g for 15 min. The supernatant was isolated, and protein was determined using the BCA protein assay kit. Brain protein extract (50 µg) was reduced and denatured and separated by electrophoresis through a 10.5 x 10 cm, 0.75-mm-thick 15% polyacrylamide preparative gel. After separation, the proteins in the gel were transferred to nitrocellulose membrane. The nitrocellulose was washed with PBS containing 0.2% Tween 20 (PBST), and remaining binding sites on the membrane were blocked by overnight incubation in PBST containing 2% nonfat dried milk at 4°C. Incubation of primary antibodies was performed at 1:300 dilution in PBST containing 2% nonfat dried milk overnight at 4°C. In some experiments, the antibodies were preincubated with 8 µg/ml of the same immunizing peptides used for the generation of the antibodies. After the nitrocellulose membrane was washed with PBST, it was incubated with an alkaline phosphatase-conjugated goat anti-rabbit secondary antibody (Sigma, St. Louis, MO), 1:2000 in PBST containing 2% nonfat dried milk for 1 hr at room temperature. The nitrocellulose membrane was washed extensively with PBST, followed by PBS. Finally, the immune complex was visualized by incubating in the presence of nitroblue tetrazolium-5-bromo-4-chloro-3-indoyl phosphate chromogen." The images can be found in Figure 1 on page 11138. Reference 2: Citation: J. Neuro. 25(10):2530-2536, 2005. Link: http://www.jneurosci.org/cgi/reprint/25/10/2530 Description: This article cites ABR's PA1-743 antibody detecting CB1 at ~85 kDa and ~52 kDa in macaque cortical tissue homogenate. "Western blotting. Protein extracts were prepared from frontal cortices of one uninfected control and two SIVE monkeys that were used for immunohistochemistry. Tissues were homogenized in lysis buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1% NP-40) with 1x protease inhibitor mixture. The homogenate was incubated on ice for 30 min and then centrifuged at 10,000 x g for 30 min at 4°C. The supernatant was collected, and protein concentration was determined using the BCA protein assay kit. Twenty-five micrograms of protein extract from each sample were reduced and denatured and separated by electrophoresis through a 4-15% gradient polyacrylamide preparative gel. The proteins were transferred from the gel to Immuno-Blot polyvinylidene difluoride membrane. The membrane was washed with TBS containing 0.1% Tween 20 (TBST) and blocked in TBST containing 5% dry goat milk. Primary antibody (ab3558) was diluted in TBST containing 5% dry goat milk at 1:2000 for CB1; this was incubated overnight at 4°C with gentle shaking. Blots were washed four times in TBST and then incubated with goat anti-rabbit HRP (1:1000) for 1 h at room temperature. The membrane was washed four times in TBST. Finally, the immune complex was visualized using an ECL Western Blotting kit. The specificity of the signal was confirmed by preincubation with the immunizing peptide (1:900)." The images can be found in Figures 1 & 2 on page 2532. With regards to ab3559 Cannabinoid Receptor I antibody, although we do not have an image available from our testing data, I have provided the original reference for this antibody. I have listed this paper as well as another reference below for your review. The paper has a wonderful western blot image on page 395 (Fig. 1 A and B) showing detection of CB1 in Sprague-Dawley rat brain extracts. References: K. Tsou et. al. Immunohistochemical Distribution of Cannabinoid CB1 Receptors in the Rat Central Nervous System. Neuroscience. 1998 Mar;83(2):393-411 Andrea L. Small-Howard et. al. Anti-inflammatory potential of CB1-mediated cAMP elevation in mast cells. Biochem. Journal 25 Jan. 2005 manuscript BJ0041682. http://www.biochemj.org/bj/imps_x/pdf/BJ20041682. I hope that this information is helpful. Please do not hesitate to contact me with any additional questions.

    Read More

    Abcam Scientific Support

    回复于 Apr 22 2008

    Question

    Thank you for your assistance. I contacted the customer and suggested her to follow your instructions. I asked her about the storage conditions and indeed, she did not notice the indication on the datasheet. However, the antibody did not entirely loose activity. Is it possible that the two very close bands she is seeing are due to transcript variants or post translational modifications? I found 2 variants in the human protein, but no clear indication in the rat case. Thank you,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 03 2007

    Answer

    Thank you for your message which has been forwarded to me on behalf of Tanya while she is away. It is with regret that we are not able to guarantee this product if it has not been stored as indicated on the datasheet. However, the results indicate that the antibody is still active and it is very likely the bands observed are from splice variants of the cannabinoid receptor 1 protein. There are splice variants of the protein in rat as well as human as indicated in the following reference: Journal of Pharmacology and Experimental Therapeutics. Vol. 282, Issue 3, 1632-1642, 1997 Regional Differences in Cannabinoid Receptor/G-protein Coupling in Rat Brain1. Christopher S. Breivogel et al. The extra band could also result from differences in post-translational modification. I hope this information is helpful. Should the customer have any further questions, please do not hesitate to contact us again.

    Read More

    Abcam Scientific Support

    回复于 Jan 03 2007

    Question

    Last week I bought rabbit anti-CB1 antibody from BioSource (Cat. 44310) and I have got the excellent result. So, I donot think there is a problem with my protocol. I wonder if I can get a refund from your company. I have spent too much time on this antibody from your company.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 18 2005

    Answer

    Thank you for your email. I'm sorry that this antibody is not working out under your conditions and I have instructed our accounting department to issue a refund for your order for ab3558. If you have any questions regarding the refund, please contact accounts@abcam.com

    Read More

    Abcam Scientific Support

    回复于 Apr 18 2005

    Question

    I used the same membrane to stain the other protein (Pten). I found this membrane works very well. This means my protocol for western blot has no problem. So, I wonder if you have this kind of antibody with different Lot No.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 05 2005

    Answer

    Thank you for your email and patience. I do suggest decreasing the concentration of the primary as well as the incubation period as the bands you are seeing may be non-specific. If you are using the same secondary with the Pten primary and not getting any additional bands then your secondary is most likely working fine. However, you probably still need to optimize the conditions with ab3558.

    Read More

    Abcam Scientific Support

    回复于 Apr 11 2005

    Question

    I have answered these questions last time. Every thing is the same. Did you test this antibody before?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 31 2005

    Answer

    Thank you for your email. Yes, this has been tested and characterized in Western blotting as well as Immunocytochemistry. In Western blotting, this antibody detects an ~60 kDa protein representing CB1 from rat brain homogenate. The bands that you are seeing at 45 and 80 kDa may be non specific and this point I would suggest decreasing the concentration of the primary as well as the incubation period. Also, make sure to run a secondary control to ensure that these bands are not due to your secondary antibody.

    Read More

    Abcam Scientific Support

    回复于 Apr 01 2005

    Question

    I haved received the replacement of CB1 antibody. I tested it and found two bands in my membrane from rat brains, one is about 80 kD, the other is about 45kD. The specific band should be ~60kD. I do not know what happened.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 30 2005

    Answer

    Thank you for your email. I recall that you were previously unable to obtain any bands with this antibody. I would like to investigate this present issue further but do need details regarding your protocol in order to do that. If you could answer the questions below, it would be very helpful. 1. Please describe the problem (high background, wrong band size, more bands, no band etc). 2. On what material are you testing the antibody in WB? •Species? •Cell extract/ Nuclear extract? •Purified protein? •Recombinant protein? 3. How much protein did you load? •How did you prepare the lysate for the analysis (protease inhibitors etc)? •Did you heat the samples? 4. Primary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? 5. Secondary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? •Do you know whether the problems you are experiencing come from the secondary? 6. What detection method are you using? 7. Background bands •Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control) •Is the blocking step sufficient? (We recommend blocking the membrane by adding 20 ml of blocking buffer (5% non-fat dry milk, 0.1% Tween-20 in TBS). Incubate for 2 h at room temperature or overnight at 4°C with agitation) •Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) •At what size are the bands migrating? Could they be degradation products of your target? •Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 8. Optimization attempts •How many times have you tried the Western? •Do you obtain the same results every time e.g. are background bands always in the same place? •What steps have you altered? 9. Did you apply positive and negative controls along with the samples? Please specify.

    Read More

    Abcam Scientific Support

    回复于 Mar 31 2005

    Question

    Thank you for your email. Have you used protease inhibitors? Also, please make sure that you are not over washing the membrane. If you have purchased the antibody in the past 90 days I can offer you a refund or replacement vial. Please let me know which you would prefer. Answer: Yes, I have used protease inhibitor (Sigma). I do not think I over washed the member. I washed the membrane just 10 min -3 times. I bought this antibody last month. I prefer a replacement vial (CB1 receptor antibody, but different lot # from my last one 94154). My Address is:

    Read More

    Abcam community

    Verified customer

    Asked on Mar 15 2005

    Answer

    There have been no previously reported problems with this particulat lot. It is a possibility that the antibody became defective during shipment. We will send you a replacement vial free of charge, please let me know how it works out for you.

    Read More

    Abcam Scientific Support

    回复于 Mar 16 2005

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    研究类别(A-Z)
    • 肿瘤研究
    • 心血管研究
    • 细胞生物学
    • 发育生物学
    • 染色质及细胞核信号转导研究
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