製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to LGR5
  • 由来種
    Rabbit
  • 特異性
    LGR5 is a multi-pass membrane protein, due to hydrophobic nature these proteins tend to aggregate when boiled at 100°C in western blotting therefore, we recommend slow heating of lysates for 15-20 minutes at 60-70°C rather than boiling. The immunogen used for this product shares 62% homology with LGR6. Cross-reactivity with this protein has not been confirmed experimentally.
  • アプリケーション
    適用あり: ICC/IF, IHC-P, Flow Cyt, WB, ELISAmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Cow
  • 免疫原

    Synthetic peptide within Human LGR5 aa 689-719 conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    Database link: O75473
    (Peptide available as ab185636)

  • ポジティブ・コントロール
    • WB: Mouse cerebellum tissue lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HepG2 cells.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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).Can be blocked with Human LGR5 peptide (ab185636).

Blocking peptide protocol

ELISA 1/1000.

ターゲット情報

  • 機能
    Orphan receptor. Stem cell marker of the intestinal epithelium and the hair follicule. Target gene of Wnt signaling.
  • 組織特異性
    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.
  • 配列類似性
    Belongs to the G-protein coupled receptor 1 family.
    Contains 16 LRR (leucine-rich) repeats.
    Contains 1 LRRNT domain.
  • 細胞内局在
    Cell membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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
    • GPCR GPR49 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

画像

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

  • All lanes : Anti-LGR5 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
    All lanes : 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 LGR5 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-LGR5 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


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

    See Abreview

  • ab75732 at 0.025 mg/ml staining LGR5 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

参考文献

This product has been referenced in:
  • Saito Y  et al. Induction of differentiation of intrahepatic cholangiocarcinoma cells to functional hepatocytes using an organoid culture system. Sci Rep 8:2821 (2018). Read more (PubMed: 29434290) »
  • Sun ZJ  et al. A fusion protein composed of the DSL domain of Dll1 and RGD motif protects cryptic stem cells in irradiation injury. Biosci Rep 38:N/A (2018). Read more (PubMed: 29444821) »

See all 19 Publications for this product

レビューと Q&A

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (adult ilium)
Permeabilization
No
Specification
adult ilium
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Methanol
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Abcam user community

Verified customer

投稿 Dec 04 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Colon and Jejunum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate acid
Permeabilization
Yes - Tween-20/Triton X 100
Specification
Colon and Jejunum
Blocking step
Casein-based blocking solution (Blocking sniper-Biocare medical) as blocking agent for 8 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Dr. Li Li

Verified customer

投稿 Mar 13 2017

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

投稿 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
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Abcam user community

Verified customer

投稿 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
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Abcam user community

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投稿 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
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投稿 Aug 20 2013

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
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Abcam user community

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投稿 Feb 12 2013

Application
Flow Cytometry
Sample
Rat Cell (Intestine)
Specification
Intestine
Preparation
Cell harvesting/tissue preparation method: 2mM EDTA digestion
Sample buffer: 1X PBS
Fixation
Paraformaldehyde
Permeabilization
No
Gating Strategy
Histogram with a negative control
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投稿 Jun 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse brain)
Loading amount
25 µg
Specification
Mouse brain
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
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Abcam user community

Verified customer

投稿 Aug 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cell (Mouse enterocytes isolated from mouse intestine)
Specification
Mouse enterocytes isolated from mouse intestine
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Aug 11 2010

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

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