Anti-LAMP2A 抗体 [EPR4207(2)] - BSA and Azide free (ab240018)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4207(2)] to LAMP2A - BSA and Azide free
- Suitable for: IHC-P, ICC/IF, Flow Cyt (Intra), IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-LAMP2A antibody [EPR4207(2)] - BSA and Azide free
LAMP2A 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR4207(2)] to LAMP2A - BSA and Azide free -
由来種
Rabbit -
特異性
LAMP2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle (PMID: 10212251PubMed:7488019, PubMed:26856698).
For better using it in tissue with low expression level, we suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate). -
アプリケーション
適用あり: IHC-P, ICC/IF, Flow Cyt (Intra), IP, WBmore details -
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
- IHC-P: Human placenta and liver tissue. Mouse and rat liver tissue. IP: HeLa and RAW 264.7 whole cell lysates. Flow Cyt (intra): HeLa cells. ICC/IF: Wild-type HeLa cells.
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特記事項
ab240018 is the carrier-free version of ab125068.
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
EPR4207(2) -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
- Alexa Fluor® 594 Anti-LAMP2A antibody [EPR4207(2)] (ab281931)
- Alexa Fluor® 647 Anti-LAMP2A antibody [EPR4207(2)] (ab284133)
- PE Anti-LAMP2A antibody [EPR4207(2)] (Lysosome Marker) (ab305387)
- APC Anti-LAMP2A antibody [EPR4207(2)] (Lysosome Marker) (ab305388)
- HRP Anti-LAMP2A antibody [EPR4207(2)] (Lysosome Marker) (ab305389)
- Alexa Fluor® 488 Anti-LAMP2A antibody [EPR4207(2)] (ab309845)
- Alexa Fluor® 568 Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab312675)
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
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Positive Controls
- HeLa whole cell lysate (ab150035)
- Mouse kidney normal tissue lysate - total protein (ab29305)
- HeLa whole cell lysate (ab29545)
- HeLa membrane extract lysate (ab29547)
- Jurkat whole cell lysate (ab30128)
- Jurkat membrane extract lysate (ab30130)
- NIH/3T3 whole cell lysate (ab7179)
- RAW 264.7 whole cell lysate (ab7187)
- Mouse kidney tissue lysate - total protein (0 days) (ab7261)
- Mouse kidney tissue lysate (14 days) (ab7262)
- Jurkat whole cell lysate (ab7899)
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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ab240018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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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
We recommend permeabilisation with 0.1% Tween-20, 5 min. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 45 kDa).
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特記事項 |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. We recommend permeabilisation with 0.1% Tween-20, 5 min. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 45 kDa). |
ターゲット情報
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機能
Implicated in tumor cell metastasis. May function in protection of the lysosomal membrane from autodigestion, maintenance of the acidic environment of the lysosome, adhesion when expressed on the cell surface (plasma membrane), and inter- and intracellular signal transduction. Protects cells from the toxic effects of methylating mutagens. -
組織特異性
Isoform LAMP-2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle. Isoform LAMP-2B is highly expressed in skeletal muscle, less in brain, placenta, lung, kidney and pancreas, very low in liver. -
関連疾患
Danon disease -
配列類似性
Belongs to the LAMP family. -
翻訳後修飾
O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans. -
細胞内局在
Cell membrane. Endosome membrane. Lysosome membrane. This protein shuttles between lysosomes, endosomes, and the plasma membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 3920 Human
- Entrez Gene: 16784 Mouse
- Omim: 309060 Human
- SwissProt: P13473 Human
- SwissProt: P17047 Mouse
- SwissProt: P17046 Rat
- Unigene: 496684 Human
- Unigene: 414016 Mouse
see all -
製品の状態
Alternative splicing produces 3 isoforms. -
別名
- CD107 antigen-like family member B antibody
- CD107b antibody
- LAMP 2 antibody
see all
画像
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All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/1000 dilution
Lane 1 : Mouse kidney tissue lysate
Lane 2 : Mouse liver tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Mouse lung tissue lysate
Lane 5 : Mouse brain tissue lysate
Lane 6 : Mouse cerebral cortex tissue lysate
Lane 7 : Mouse heart tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 40 secondsThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
Blocking and diluting buffer and concentration: 5% NFDM /TBST.
ab181602 was used as a GAPDH loading control.
LAMP2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle (PMID: 10212251PubMed:7488019, PubMed:26856698).
For better using it in tissue with low expression level, we suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate). -
Side-by-side comparison of ICC performance using the rabbit polyclonal ab18528 and RabMab® ab125068. Staining was performed on wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% Tween-20 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 ab18528 or ab125068 overnight at +4°C at 3 different concentrations: 1.0 µg/mL, 0.2 µg/mL and 0.04 µg/mL. Secondary antibody incubation was at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) at 1/1000 and nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Some cytoplasmic cross-reactivity is seen using ab18528 at 1.0 µg/mL, but further titration of the antibody improves the ICC staining result. The RabMab® ab125068 shows negligible non-specific staining across the dilution range. Quantification of the antibody signal was performed using a minimum of 135 cells and data are presented as mean ± SD.
Optimal dilutions/concentrations may vary across different cell types/experiment conditions and should be determined by the end user.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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Side-by-side comparison of ICC performance using the rabbit polyclonal ab18528 and RabMab® ab125068. Staining was performed on wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Tween-20 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 ab18528 or ab125068 overnight at +4°C at 3 different concentrations: 1.0 µg/mL, 0.2 µg/mL and 0.04 µg/mL. Secondary antibody incubation was at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) at 1/1000 and nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Some cytoplasmic cross-reactivity is seen using ab18528 at 1.0 µg/mL, but further titration of the antibody improves the ICC staining result. The RabMab® ab125068 shows negligible non-specific staining across the dilution range. Quantification of the antibody signal was performed using a minimum of 180 cells and data are presented as mean ± SD.
Optimal dilutions/concentrations may vary across different cell types/experiment conditions and should be determined by the end user.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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ab125068 staining LAMP2a in wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Tween-20 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 ab125068 at 0.04 µg/mL and ab7291 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) and goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) (shown in red) both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
In our hands, permeabilization with 0.1% Triton X-100 (5 min) resulted in greatly reduced signal and we recommend using 0.1% Tween-20 (5 min) for detecting this target.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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ab125068 staining LAMP2a in wild-type HeLa cells (top panel) and LAMP2 knockout HeLa cells (bottom panel, available as ab255402). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Tween-20 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 ab125068 at 0.04 µg/mL and ab7291 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in green) and goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) (shown in red) both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
In our hands, permeabilization with 0.1% Triton X-100 (5 min) resulted in greatly reduced signal and we recommend using 0.1% Tween-20 (5 min) for detecting this target.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling LAMP2A (red) with ab125068 at a 1/1000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling LAMP2A with purified ab125068 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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ab125068 (purified) at 1/60 immunoprecipitating LAMP2A in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab125068 + HeLa whole cell lysate (10µg).
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab125068 in HeLa whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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LAMP2A was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate with ab125068 at 1/100 dilution. Western blot was performed from the immunoprecipitate using unpurified ab125068 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: RAW 264.7 whole cell lysate 10ug (Input).
Lane 2: ab125068 IP in RAW 264.7 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab125068 in RAW 264.7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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LAMP2A was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with unpurified ab125068 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab125068 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate 10ug (Input).
Lane 2: ab125068 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab125068 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
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Immunohistochemical analysis of paraffin-embedded Rat liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on rat liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on mouse liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded Human liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on human liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded Human placenta labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on human placenta tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125068).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (0)
ab240018 は論文での使用が確認できていません。