概述

  • 产品名称Anti-GPCR GPR49抗体
    参阅全部 GPCR GPR49 一抗
  • 描述
    兔多克隆抗体to GPCR GPR49
  • 经测试应用适用于: ICC/IF, IHC-P, Flow Cyt, WB, ELISAmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Cow
  • 免疫原

    Synthetic peptide corresponding to Human GPCR GPR49 aa 689-719 conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab185636)

  • 阳性对照
    • WB: Mouse cerebellum tissue lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HepG2 cells.

性能

应用

Our Abpromise guarantee covers the use of ab75732 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/100.
IHC-P 1/10 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Please note for better results the antigen retrieval method may need optimizations. We recommend trying a range of incubation time e.g. 1, 2, 3, 4, 5 minutes using pressure cooker or 3, 5, 10, 15 minutes in microwave after buffer start boiling. For more details check IHC-P – detailed guide in protocol section.

Flow Cyt 1/10 - 1/50. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 100 kDa).

Blocking peptide protcol

ELISA 1/1000.

靶标

  • 功能Orphan receptor. Stem cell marker of the intestinal epithelium and the hair follicule. Target gene of Wnt signaling.
  • 组织特异性Expressed in skeletal muscle, placenta, spinal cord, and various region of brain. Expressed at the base of crypts in colonic and small mucosa stem cells. In premalignant cancer expression is not restricted to the cript base. Overexpressed in cancers of the ovary, colon and liver.
  • 序列相似性Belongs to the G-protein coupled receptor 1 family.
    Contains 16 LRR (leucine-rich) repeats.
    Contains 1 LRRNT domain.
  • 细胞定位Cell membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • FEX antibody
    • G protein coupled receptor 49 antibody
    • G protein coupled receptor 67 antibody
    • g protein-coupled receptor fex antibody
    • G-protein coupled receptor 49 antibody
    • G-protein coupled receptor 67 antibody
    • G-protein coupled receptor HG38 antibody
    • GPR 49 antibody
    • GPR 67 antibody
    • GPR49 antibody
    • GPR67 antibody
    • GRP 49 antibody
    • GRP49 antibody
    • HG 38 antibody
    • HG38 antibody
    • Leucine rich repeat containing G protein coupled receptor 5 antibody
    • Leucine-rich repeat-containing G-protein coupled receptor 5 antibody
    • LGR 5 antibody
    • LGR5 antibody
    • LGR5_HUMAN antibody
    • MGC117008 antibody
    • Orphan G protein coupled receptor HG38 antibody
    see all

Anti-GPCR GPR49 antibody 图像

  • Immunohistochemical analysis of PFA fixed, paraffin-embedded human brain tissue sections labelling LGR5 with ab75732 at 1/25 dilution.

  • All lanes : Anti-GPCR GPR49 antibody (ab75732) at 1/2000 dilution

    Lane 1 : Mouse skeletal muscle lysates
    Lane 2 : Rat skeletal muscle lysates
    Lane 3 : Hela whole cell lysates
    Lane 4 : HepG2 whole cell lysates
    Lane 5 : SH-SY5Y whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution

    Predicted band size : 100 kDa

    Blocking buffer 5% NFDM/TBST

    Diluting buffer 5% NFDM/TBST

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling GPCR GPR49 with ab75732. Cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min) and then incubated with the primary antibody (1/100, 2 h at room temperature). An Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used as the secondary antibody (1/1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min).

  • Overlay histogram showing 2% paraformaldehyde fixed SH-SY5Y cells stained with ab75732 (green line). Cells were incubated with the antibody at 1/25 dilution at 37oC for 60 minutes. The secondary antibody used was DyLight® 488 Goat-Anti-Rabbit IgG at 1/400 dilution. A non-specific IgG antibody was used for an isotype control (blue line).

  • Immunohistochemical analysis of PFA fixed, paraffin-embedded mouse brain tissue sections labelling LGR5 with ab75732 at 1/25 dilution.

  • Anti-GPCR GPR49 antibody (ab75732) at 1/750 dilution (for 16 hours at 4°C) + Whole tissue lysate of mouse brain at 18 µg

    Secondary
    An HRP-conjugated Goat polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 100 kDa
    Observed band size : 97 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    Blocking Step: 5% Milk for 1 hour at 20°C.

    See Abreview

  • ab75732 at 0.025 mg/ml staining GPCR GPR49 in human Hela cells by Immunocytochemistry / Immunofluorescence.

  • Overlay histogram showing mouse intestinal cells stained with ab75732 (pink line). Cells were incubated with the antibody at 1/50 in DMEM at 20°C for 30 minutes.The secondary antibody used was DyLight® 649 goat anti-rabbit IgG at 1/100 dilution. A non-specific IgG antibody was used for an isotype control (black line).

    See Abreview

Anti-GPCR GPR49 antibody (ab75732)参考文献

This product has been referenced in:
  • Sun G  et al. Inhibitory effects of Hedyotis diffusa Willd. on colorectal cancer stem cells. Oncol Lett 11:3875-3881 (2016). WB ; Human . Read more (PubMed: 27313710) »
  • Kitamura J  et al. Chronic lung injury by constitutive expression of activation-induced cytidine deaminase leads to focal mucous cell metaplasia and cancer. PLoS One 10:e0117986 (2015). IHC ; Mouse . Read more (PubMed: 25659078) »

See all 10 Publications for this product

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Thanks for your reply.

I am sorry to hear some of your colleagues have had a bad experience regarding our "Abpromise". I would encourage them to contact us to resolve any issues they have.

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Thank you for contacting us. Below is the protocol that was used: Preparing Tissue Sections for Immunostaining: • Fix the tissue in 10% formalin at 4°C overnight. • Embed fixed tissue in paraffin. • Mount tissue sections on slides. • Clear t...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization No
Specification Colon
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
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Verified customer

提交于 Oct 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Bos taurus Tissue sections (Airway)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
Permeabilization No
Specification Airway
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 23 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step None
Sample Human Tissue sections (tooth)
Specification tooth
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 07 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Fixation none
Permeabilization No
Sample Mouse Cell (intestinal cells)
Specification intestinal cells
Gating Strategy high FSC/SSC popn log scale
Preparation Cell harvesting/tissue preparation method: isolate colon, collagenase digestion
Sample buffer: DMEM
Username

Abcam user community

Verified customer

提交于 Aug 20 2013

While we do not have specific data regarding this products use in FACS sorting, the immunogen area is within an extracellular region. As such, it seems very likely that this antibody will be successful in that application.

Thank you for the clarification.
You are right, this antibody is indeed tested in Flow, but with a conjugated secondary antibody.
If you would like to use this antibody conjugated directly to a fluorochrome, I would suggest to have a look at ...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (intestine)
Specification intestine
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Citrate
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 12 2013

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact ...

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