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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microtubules MT Associated Proteins MAP
使用敲除细胞株进行验证重组RabMAb

重组Anti-LC3B抗体[EPR18709] - Autophagosome Marker (ab192890)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (15) Submit a question References (82)

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Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
  • Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18709] to LC3B - Autophagosome Marker
  • Suitable for: IP, WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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概述

  • 产品名称

    Anti-LC3B抗体[EPR18709] - Autophagosome Marker
    参阅全部 LC3B 一抗
  • 描述

    兔单克隆抗体[EPR18709] to LC3B - Autophagosome Marker
  • 宿主

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: BMDM, U-87 MG, C6 and RAW 264.7 whole cell lysates; Human brain, mouse heart, rat heart, mouse brain and rat brain lysates. IP: HeLa whole cell lysate. ICC/IF: HeLa cells (+/- chloroquine), HAP1 cells (+/- chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line).
  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 long term. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EPR18709
  • 同种型

    IgG
  • 研究领域

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • MT Associated Proteins
    • MAP
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Cancer
    • Signal transduction
    • Autophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • APG gene products
    • Cancer
    • Cell Death
    • Autophagy
    • Signal Transduction
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products

相关产品

  • Alternative Versions

    • Anti-LC3B antibody [EPR18709] - BSA and Azide free (ab221794)
    • Alexa Fluor® 488 Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab225382)
    • Alexa Fluor® 647 Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab225383)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human LC3B protein (ab103506)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab192890 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
ICC/IF
Human
IP
Human
WB
Mouse
Rat
Human
应用 Ab评论 说明
IP
1/50.
WB (8)
1/2000. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa).
ICC/IF (4)
Use a concentration of 1 µg/ml.
说明
IP
1/50.
WB
1/2000. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa).
ICC/IF
Use a concentration of 1 µg/ml.

靶标

  • 功能

    Probably involved in formation of autophagosomal vacuoles (autophagosomes).
  • 组织特异性

    Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
  • 序列相似性

    Belongs to the MAP1 LC3 family.
  • 翻译后修饰

    The precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II.
  • 细胞定位

    Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
  • Target information above from: UniProt accession Q9GZQ8 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: 81631 Human
    • Entrez Gene: 67443 Mouse
    • Entrez Gene: 64862 Rat
    • Omim: 609604 Human
    • SwissProt: Q9GZQ8 Human
    • SwissProt: Q9CQV6 Mouse
    • SwissProt: Q62625 Rat
    • Unigene: 356061 Human
    • Unigene: 28357 Mouse
    • Unigene: 41412 Rat
    see all
  • 别名

    • ATG8F antibody
    • Autophagy-related protein LC3 B antibody
    • Autophagy-related ubiquitin-like modifier LC3 B antibody
    • LC3B antibody
    • LC3II antibody
    • MAP1 light chain 3 like protein 2 antibody
    • MAP1 light chain 3-like protein 2 antibody
    • MAP1A/1BLC3 antibody
    • MAP1A/MAP1B LC3 B antibody
    • MAP1A/MAP1B light chain 3 B antibody
    • MAP1ALC3 antibody
    • MAP1LC3B a antibody
    • Map1lc3b antibody
    • Microtubule associated protein 1 light chain 3 beta antibody
    • Microtubule-associated protein 1 light chain 3 beta antibody
    • Microtubule-associated proteins 1A/1B light chain 3B antibody
    • MLP3B_HUMAN antibody
    see all

图片

  • Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LC3B knockout HAP1 cell lysate (20 µg)
    Lane 3: Human brain tissue lysate (20 µg)
    Lane 4: U-87 MG cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green).

    Green -target observed at 14 and 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab192890 and a competitor's top cited rabbit polyclonal antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

    ab192890 staining LC3B in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Chloroquine (50μM, 24 hours).

    The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Alexa Fluor® 488 (ab225383) and Alexa Fluor® 647 (ab225382) conjugated versions are available for this clone.

  • Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Immunoprecipitation - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

    LC3B was immunoprecipitated from 1 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab192890 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab192890 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
    Lane 1: HeLa whole cell lysate 10 µg (Input).
    Lane 2: ab192890 IP in HeLa whole cell lysate.
    Lane 3: Rabbit IgG,monoclonal [EPR25A] Isotype Control (ab172730) instead of ab192890 in HeLa whole cell cell lysate.
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 30 seconds.

  • Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

    ab192890 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50μM, 24 hours).

    The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192890 at 1 μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LC3B knockout HAP1 cell lysate (20 µg)
    Lane 3: Human brain tissue lysate (20 µg)
    Lane 4: U-87 MG cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab192890 observed at 14 and 16 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab192890 was shown to specifically react with LC3B in wild-type HAP1 cells. No band was observed when LC3B knockout samples were examined. Wild-type and LC3B knockout samples were subjected to SDS-PAGE. ab192890 and ab8245 (loading control to GAPDH) were diluted 1/2000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Western blot - Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    All lanes : Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) at 1/2000 dilution

    Lane 1 : Human brain lysate
    Lane 2 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : Mouse heart lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Mouse brain lysate
    Lane 8 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 15 kDa
    Observed band size: 14,16 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1-6: 3 minutes; Lane 7 and 8: 30 seconds.

  • Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
    Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)

实验方案

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • 文献 (82)

    发表研究结果有使用 ab192890?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab192890 被引用在 82 文献中.

    • Cassidy LD  et al. Temporal inhibition of autophagy reveals segmental reversal of ageing with increased cancer risk. Nat Commun 11:307 (2020). WB ; Mouse . PubMed: 31949142
    • Khorraminejad-Shirazi M  et al. AICAR and nicotinamide treatment synergistically augment the proliferation and attenuate senescence-associated changes in mesenchymal stromal cells. Stem Cell Res Ther 11:45 (2020). PubMed: 32014016
    • Sasaki N  et al. Rapamycin promotes endothelial-mesenchymal transition during stress-induced premature senescence through the activation of autophagy. Cell Commun Signal 18:43 (2020). PubMed: 32164764
    • Kong Q  et al. Autophagy inhibits TLR4-mediated invasiveness of oral cancer cells via the NF-?B pathway. Oral Dis N/A:N/A (2020). PubMed: 32291890
    • Liu H  et al. Overexpression of the clock gene Per2 suppresses oral squamous cell carcinoma progression by activating autophagy via the PI3K/AKT/mTOR pathway. J Cancer 11:3655-3666 (2020). PubMed: 32284762
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Glioblastoma cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    Glioblastoma cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    提交于 Dec 08 2020

    Western blot abreview for Anti-LC3B antibody [EPR18709]

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa and RPE-1)
    Gel Running Conditions
    Reduced Denaturing (12% gel)
    Loading amount
    100000 cells
    Treatment
    HBSS starvation for 2h
    Specification
    HeLa and RPE-1
    Blocking step
    Milk as blocking agent for 45 minute(s) · Concentration: 5%
    Read More

    Dr. Marie Macurkova

    Verified customer

    提交于 Mar 28 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3B antibody [EPR18709]

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    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Primary human skin-derived fibroblasts)
    Permeabilization
    Yes - Triton x100 0.1%
    Specification
    Primary human skin-derived fibroblasts
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Trine Mogensen

    Verified customer

    提交于 Feb 20 2019

    Western blot abreview for Anti-LC3B antibody [EPR18709]

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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human skin-derived fibroblasts, primary)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    3 µg
    Treatment
    Herpes simplex virus-2
    Specification
    Human skin-derived fibroblasts, primary
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Dr. Trine Mogensen

    Verified customer

    提交于 Feb 19 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3B antibody [EPR18709]

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    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Cardiomyocytes)
    Permeabilization
    Yes - Triton x-100, 0.03%
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
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    提交于 Dec 15 2018

    Western blot abreview for Anti-LC3B antibody [EPR18709]

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    Application
    Western blot
    Sample
    Monkey Cell lysate - whole cell (Heart protein lysate (Rhesus))
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    Heart protein lysate (Rhesus)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

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    提交于 Nov 30 2018

    Western blot abreview for Anti-LC3B antibody [EPR18709]

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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEk293)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    HEk293
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

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    提交于 Nov 30 2018

    Immunocytochemistry abreview for Anti-LC3B antibody [EPR18709]

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    Application
    Immunocytochemistry
    Sample
    Rat Cultured Cells (H9C2 rat cardiomyocytes)
    Permeabilization
    Yes - Triton x-100, 0.03%
    Specification
    H9C2 rat cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
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    提交于 Nov 13 2018

    Western blot abreview for Anti-LC3B antibody [EPR18709]

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Kidney (HEK293 cells))
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    30 µg
    Specification
    Kidney (HEK293 cells)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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    提交于 Feb 15 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-LC3B antibody [EPR18709]

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    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    African green monkey Cell (Kidney)
    Permeabilization
    Yes - 0.1% triton x-100
    Specification
    Kidney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Methanol
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    研究类别(A-Z)
    • 肿瘤研究
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