Anti-LC3B 抗体 [EPR18709] - BSA and Azide free (ab221794)
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 - BSA and Azide free
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-LC3B antibody [EPR18709] - BSA and Azide free
LC3B 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR18709] to LC3B - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IFmore details
適用なし: IP -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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. ICC/IF: HeLa cells (+/- chloroquine), HAP1 cells (+/- chloroquine) (HAP1-MAP1LC3B knockout cells used as negative cell line); H9C2 rat cardiomyocytes. IHC-P: Human Cerebral Cortex tissue sections
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特記事項
ab221794 is the carrier-free version of ab192890.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR18709 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890)
- Alexa Fluor® 488 Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab225382)
- Alexa Fluor® 647 Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab225383)
- Autophagy Analysis (ATG16L1, ATG16L1 pS278, SQSTM1, LC3B, Ubiquitin, M6PR) Antibody Sampler Panel (ab269811)
- Alexa Fluor® 555 Anti-LC3B - Autophagosome Marker antibody [EPR18709] (ab307768)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab221794の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa).
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IHC-P |
Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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特記事項 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 14, 16 kDa (predicted molecular weight: 15 kDa). |
IHC-P
Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
ターゲット情報
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機能
Probably involved in formation of autophagosomal vacuoles (autophagosomes). -
組織特異性
Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver. -
配列類似性
Belongs to the MAP1 LC3 family. -
翻訳後修飾
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. -
細胞内局在
Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes. - Information by UniProt
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参照データベース
- 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
see all -
別名
- ATG8F antibody
- Autophagy-related protein LC3 B antibody
- Autophagy-related ubiquitin-like modifier LC3 B antibody
see all
画像
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All lanes : Anti-LC3B antibody [EPR18709] - Autophagosome Marker (ab192890) at 1/2000 dilution
Lane 1 : Wild-type HepG2 untreated control cell lysate
Lane 2 : Wild-type HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 3 : MAP1LC3B knockout HepG2 untreated control cell lysate
Lane 4 : MAP1LC3B knockout HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 5 : U-87 MG cell lysate
Lane 6 : PC-12 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 14,16 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
False colour image of Western blot: Anti-LC3B antibody [EPR18709] - Autophagosome Marker staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab192890 was shown to bind specifically to LC3B. A band was observed at 16/14 kDa (yellow arrows) in treated wild-type HepG2 cell lysates with no signal observed at this size in MAP1LC3B knockout cell line ab277828 (knockout cell lysate ab283796). To generate this image, wild-type and MAP1LC3B knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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ab192890 staining LC3B in HeLa 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
ab192890 staining LC3B in paraffin embedded human astrocytoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Samples were incubated with primary antibody at 1/1000 dilution for 30 mins at room temperature. Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
Immunohistochemical analysis of formalin fixed paraffin embedded human cortex labelling LC3B with ab192890 at a dilution of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5) . ab192890 anti LC3B antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
Tissue Microarrays stained for Anti-LC3B antibody [EPR18709] - Autophagosome Marker using ab192890 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested.
Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.
The section was incubated with ab192890 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
IHC image of LC3B staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab221794, 0.1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Clone EPR18709 (ab221794) has been successfully conjugated by Abcam. This image was generated using Anti-LC3B antibody [EPR18709] - Autophagosome Marker (Alexa Fluor® 647). Please refer to ab225383 for protocol details.
ab225383 staining LC3B in wild-type HAP1 cells and knockout cells, untreated and chloroquine-treated (50μM, 24 hours). The cells were fixed with 100% methanol (5min), 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 with ab225383 at 0.5μg/ml (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone EPR18709 (ab221794) has been successfully conjugated by Abcam. This image was generated using Anti-LC3B antibody [EPR18709] - Autophagosome Marker (Alexa Fluor® 488). Please refer to ab225382 for protocol details.
ab225382 staining LC3B in HeLa chloroquine-treated (50µM, 24 hours) cells. 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 ab225453 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling LC3B with ab192890 at 1/500 dilution. Cells were fixed in Methanol. Staining with ab192890 at 1/500 was carried out for 1 hour at 22°C in PBS buffer. ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody was used at 1/200 dilution. DAPI was used to counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192890).
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This ICC data was generated using the same anti-LC3B antibody clone, EPR18709, in a different buffer formulation (cat# 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 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
プロトコール
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (5)
ab221794 は 5 報の論文で使用されています。
- Zhang H et al. Circ_0020123 plays an oncogenic role in non-small cell lung cancer depending on the regulation of miR-512-3p/CORO1C. Thorac Cancer 13:1406-1418 (2022). PubMed: 35388975
- Yu L et al. Blockage of AMPK-ULK1 pathway mediated autophagy promotes cell apoptosis to increase doxorubicin sensitivity in breast cancer (BC) cells: an in vitro study. BMC Cancer 21:195 (2021). PubMed: 33632157
- Li XR et al. Knockdown of FBP1 enhances radiosensitivity in prostate cancer cells by activating autophagy. Neoplasma 67:982-991 (2020). PubMed: 32453596
- Chen Q et al. Knockdown of Parkinson's disease-related gene ATP13A2 reduces tumorigenesis via blocking autophagic flux in colon cancer. Cell Biosci 10:144 (2020). PubMed: 33308286
- Kang C et al. Baicalin alleviates 6-hydroxydopamine-induced neurotoxicity in PC12 cells by down-regulation of microRNA-192-5p. Brain Res N/A:N/A (2018). PubMed: 30552896