製品の概要

  • 製品名
    Anti-LAMP1 antibody - Lysosome Marker
    LAMP1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to LAMP1 - Lysosome Marker
  • アプリケーション
    適用あり: IP, IHC-P, ICC, ICC/IF, WB, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Hamster, Cat, Dog, Human, Xenopus laevis, Zebrafish, African green monkey
    交差が予測される動物種: Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human LAMP1.

    (Peptide available as ab25744.)

  • ポジティブ・コントロール
    • WB: Jurkat (Human T cell lymphoblast-like cell line), A431 (Human epithelial carcinoma cell line), HEK293 (Human embryonic kidney cell line) and MCF7 (Human breast adenocarcinoma cell line) whole cell lysates.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab24170 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
IP Use at an assay dependent concentration. PubMed: 21152024
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.

We recommend to use PFA fixation as fixation with 100% Methanol can lead to staining artefacts.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 90-120 kDa (predicted molecular weight: 120 kDa).

Abcam recommends using Milk as the blocking agent.

IHC-Fr Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Presents carbohydrate ligands to selectins. Also implicated in tumor cell metastasis.
  • 配列類似性
    Belongs to the LAMP family.
  • 翻訳後修飾
    O- and N-glycosylated; some of the 18 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
  • 参照データベース
  • 別名
    • CD107 antigen like family member A antibody
    • CD107 antigen-like family member A antibody
    • CD107a antibody
    • CD107a antigen antibody
    • LAMP 1 antibody
    • LAMP-1 antibody
    • LAMP1 antibody
    • LAMP1_HUMAN antibody
    • LAMPA antibody
    • LGP120 antibody
    • lgpA antibody
    • Lysosomal membrane glycoprotein 120KD antibody
    • Lysosomal Associated Membrane Protein 1 antibody
    • Lysosome associated membrane glycoprotein 1 antibody
    • Lysosome-associated membrane glycoprotein 1 antibody
    • Lysosome-associated membrane protein 1 antibody
    • OTTHUMP00000040663 antibody
    see all

画像

  • All lanes : Anti-LAMP1 antibody - Lysosome Marker (ab24170) at 1 µg/ml

    Lane 1 : Jurkat (Human) Whole Cell Lysate
    Lane 2 : HEK293 (Human) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 120 kDa
    Additional bands at : 20 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab24170 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • ab24170 staining LAMP1 in human kidney tissue sections. Staining correlates with lysosomal specificity, particularly in the proximal convoluted tubules where lysosomes are enriched. Formalin/PFA-fixed human kidney tissue sections were incubated with ab24170 (1/200) for 2 hours. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Please see accompanying abreview for additional information.

    See Abreview

  • ICC/IF image of ab24170 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab24170, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • All lanes : Anti-LAMP1 antibody - Lysosome Marker (ab24170) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 120 kDa
    Additional bands at : 23 kDa,35 kDa,45 kDa. We are unsure as to the identity of these extra bands.
  • NOTE: The filamentous pattern observed only occurs when cell are fixed with 100% methanol. We are aware that the staining pattern can change if the cells are fixed with 4% PFA or an alternative fixative. LAMP-1 is a lysosome marker but also plays a role in cellular adhesion. Therefore, lysosomal and filamentous patterns for LAMP-1 can be possible.

    ab24170 staining Lamp-1 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab24170 at 1μg/ml and ab7291 (staining Tubulin) at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Alexa Fluor® 488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green) and Alexa Fluor® 594 Goat anti-Mouse (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI, which was added to the secondary antibody mixture.

  • IHC-P image of LAMP1 staining on human Cortex sections using ab24170 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retreival using citric acid. The sections were then permeabilized using 0.05% Tween-20 and blocking was performed using 3% BSA for 1 hour at 21°C. The primary antibody ab24170 was diluted using  3% BSA with 0.05% Tween-20 in PBS and incubated  with the sections for 18 hours at 4°C. The secondary antibody used was Goat polyclonal to rabbit IgG conjugated to biotin (1:500)

    See Abreview

  • Anti-LAMP1 antibody - Lysosome Marker (ab24170) at 1/700 dilution (in 5% milk for 4 hours at 20°C) + Rat Kidney - whole tissue lysate at 18 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 120 kDa


    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

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

    See Abreview

  • ab24170 staining LAMP1 in human fibrosarcoma cells by Immunocytochemistry/ Immunofluorescence. The cells were PFA fixed, permeabilised in 0.1% Triton X-100 and incubated with the primary antibody at 1µg/ml for 1 hour at 20°C. The secondary antibody used was a donkey anti-rabbit (H+L) IgG conjugated to Alexa Fluor® 555 used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).

    See Abreview

  • ab24170 staining LAMP1 in Human Retinal Microvascular Endothelial Cells (HRMEC) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton in PBS and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/300 in PBS) for 1 hour at 22°C. A diluted (1/500) Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Immunocytochemical immunofluorescence analyis of PFA-fixed rat primary alveolar type II cells, labelling LAMP1 with ab24170 at a dilution of 1/200 incubated for 1/200. Blocking was with 10% BSA incubated for 5 minutes. Secondary was a donkey anti-rabbit polyclonal Alexa Fluor® 647 at 1/400.

    See Abreview

  • ICC/IF image of ab24170 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24170, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

参考文献

This product has been referenced in:
  • Fujihira H  et al. Lethality of mice bearing a knockout of the Ngly1-gene is partially rescued by the additional deletion of the Engase gene. PLoS Genet 13:e1006696 (2017). WB ; Mouse . Read more (PubMed: 28426790) »
  • Sambri I  et al. Lysosomal dysfunction disrupts presynaptic maintenance and restoration of presynaptic function prevents neurodegeneration in lysosomal storage diseases. EMBO Mol Med 9:112-132 (2017). Read more (PubMed: 27881461) »

See all 178 Publications for this product

レビューと Q&A

Filter by Application

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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投稿 Nov 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (fat)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
fat
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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投稿 Nov 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
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投稿 Nov 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
skin tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin
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投稿 Nov 06 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cow Cell lysate - whole cell (Lung)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Lung
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 11 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 11 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 11 2017

Application
Western blot
Sample
African green monkey Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 10 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (kidney)
Permeabilization
Yes - 0.2% triton x100
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde
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投稿 Mar 24 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
African green monkey Cell (kidney)
Permeabilization
Yes - 0.2% triton x100
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
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投稿 Mar 24 2017

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