概述

  • 产品名称Anti-LAMP2抗体[H4B4]
    参阅全部 LAMP2 一抗
  • 描述
    小鼠单克隆抗体[H4B4] to LAMP2
  • 特异性Human CD107b/LAMP-2
  • 经测试应用适用于: Flow Cyt, IHC-Fr, ICC, WB, IHC-P, ICC/IF, IHC-FoFrmore details
  • 种属反应性
    与反应: Human, Rhesus monkey, African Green Monkey
    不与反应: Mouse, Rat
  • 免疫原

    The details of the immunogen for this antibody are not available.

应用

Our Abpromise guarantee covers the use of ab25631 in the following tested applications.

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

应用 Ab评论 说明
Flow Cyt Use 0.5-1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

In our hands milk blocking is gives superior results to BSA blocking for this product.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100.
IHC-FoFr Use at an assay dependent concentration. (PMID 19837699).

靶标

  • 功能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.
  • 组织特异性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.
  • 疾病相关Defects in LAMP2 are the cause of Danon disease (DAND) [MIM:300257]; also known as glycogen storage disease type 2B (GSD2B). DAND is a lysosomal glycogen storage disease characterized by the clinical triad of cardiomyopathy, vacuolar myopathy and mental retardation. It is often associated with an accumulation of glycogen in muscle and lysosomes.
  • 序列相似性Belongs to the LAMP family.
  • 翻译后修饰O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.
  • 细胞定位Cell membrane. Endosome membrane. Lysosome membrane. This protein shuttles between lysosomes, endosomes, and the plasma membrane.
  • Information by UniProt
  • 数据库链接
  • 形式Alternative splicing produces 3 isoforms.
  • 别名
    • CD107 antigen-like family member B antibody
    • CD107b antibody
    • LAMP 2 antibody
    • LAMP 2C antibody
    • LAMP-2 antibody
    • LAMP2 antibody
    • LAMP2_HUMAN antibody
    • LAMPB antibody
    • LGP110 antibody
    • Lysosomal associated membrane protein 2 antibody
    • Lysosome associated membrane protein 2 antibody
    • Lysosome-associated membrane glycoprotein 2 antibody
    • Lysosome-associated membrane protein 2 antibody
    • MAC3 antibody
    see all

Anti-LAMP2 antibody [H4B4] 图像

  • All lanes : Anti-LAMP2 antibody [H4B4] (ab25631) at 1/500 dilution

    Lane 1 : HEK293 cell lysate at 30 µg
    Lane 2 : HepG2 at 30 µg
    Lane 3 : THP-1 at 30 µg
    Lane 4 : 3T3-L1 at 30 µl
    Lane 5 : Mouse hepatocytes at 30 µl
    Lane 6 : Rat hepatocytes at 30 µl

    Secondary
    HRP conjugated Goat anti-Mouse IgG at 1/2000 dilution
    Developed using the ECL technique

    Exposure time : 30 seconds

    This image is of an Abreciew submitted by Daniel Rodriguez.

    Along with THP-1 macrophages, other human cell lines were loaded, and the 110 kDa mature band for LAMP2 was detected in all the samples. On the other hand, mouse and rat cells were negative. The antibody works really good on human samples, detecting a single 110 kDa band but it's not suitable to use for mouse or rat samples.

    See Abreview

  • All lanes : Anti-LAMP2 antibody [H4B4] (ab25631) at 1/500 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : Human liver lysate
    Lane 4 : Human liver membrane fraction lysate
    Lane 5 : Human skeletal muscle lysate
    Lane 6 : Human brain lysate
    Lane 7 : Human kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

    Performed under reducing conditions.

    Additional bands at : 100 kDa (possible post-translational modification),110 kDa (possible post-translational modification),120 kDa (possible post-translational modification).

    Exposure time : 20 minutes

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes.  After transfer, the membrane was blocked for an hour in 3% milk before being incubated overnight at 4°C with mouse monoclonal [H4B4] to LAMP2 (ab25631; diluted 1:5000).  Antibody binding was detected using peroxidase labelled goat anti-mouse IgG (ab97040; diluted 1:50000) for an hour at room temperature and visualised using ECL development solution.

  • ab25631 at 0.5µg/ml staining human HEK cells by immunocytochemistry. The cells were fixed with paraformaldehyde and incubated with the antibody for 1 hour. Bound antibody was detected using a goat anti-mouse IgG Alexa-Fluor ® 568. In the confocal image ab25631 labelling in red shows a distribution consistent with the location of lysosomes. Blue nuclear counterstain is present.

    This image is courtesy of an Abreview submitted by Randal Moldrich on 31 March 2006.

    See Abreview

  • ab25631 staining LAMP2 in human THP-1 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 and blocking with 1% BSA at 220C for 2 hours was done. Samples were incubated with primary antibody (1/400: in PBST and 1% BSA) for 16 hour at 40C. An Alexa Fluor ® 568-conjugated goat polyclonal to mouse IgG was used at dilution at 1/500 as secondary antibody. Merge figure shows the bright field image and fluorescent image combined.

    See Abreview

  • Anti-LAMP2 antibody [H4B4] (ab25631) at 1/500 dilution + whole cell lysate prepared from THP-1 macrophages at 30 µg

    Secondary
    Goat anti-mouse IgG conjugated to HRP at 1/2000 dilution
    Developed using the ECL technique

    Observed band size : 110 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image was kindly supplied by Daniel Rodriguez by Abreview

    Primary antibody incubated for 12 hours at 4°C.
    Gel running conditions: 12%
    Blocked with 5% milk for 1 hour at 25°C.

    See Abreview



  • Performed under reducing conditions.

    Observed band size : 110 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 second

    This image is courtesy of an anonymous Abreview

    Western blot analysis of Rhesus monkey primary retinal pigmented epthelium whole cell lysate and HeLa cell whole cell lysate (20µg/lane) labelling LAMP2 with ab25631 at 1/1000. An alkaline phosphatase-conjugated rabbit anti-mouse IgG was used as the secondary antibody.

    See Abreview

  • Flow Cytometry analysis of Human T lymphocyte cell line Jurkat labeling LAMP2 with ab25631 at 1 μg/106 cells dilution (purple). A Goat Anti-Mouse IgG1, Human ads-FITC was used as the secondary antibody. Grey - Isotype Control, Mouse IgG1-UNLB, followed by Goat Anti-Mouse IgG1, Human ads-FITC.

  • Ab25631 staining human normal placenta. Staining is localized to the cytoplasm.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Overlay histogram showing THP1 cells stained with ab25631 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab25631, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-LAMP2 antibody [H4B4] (ab25631)参考文献

This product has been referenced in:
  • Jung J  et al. Amino Acid-Dependent mTORC1 Regulation by the Lysosomal Membrane Protein SLC38A9. Mol Cell Biol 35:2479-94 (2015). Read more (PubMed: 25963655) »
  • Ghosh A  et al. Activation of peroxisome proliferator-activated receptor a induces lysosomal biogenesis in brain cells: implications for lysosomal storage disorders. J Biol Chem 290:10309-24 (2015). Read more (PubMed: 25750174) »

See all 31 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12.5)
Sample Rhesus monkey Cell lysate - whole cell (Rhesus monkey primary Retinal pigmented epithelium)
Specification Rhesus monkey primary Retinal pigmented epithelium
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
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提交于 May 13 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Skeletal muscle from the gastrocnemius)
Specification Skeletal muscle from the gastrocnemius
Fixative Methacarn
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 23°C
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Verified customer

提交于 Dec 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - other (Skeletal Muscle)
Loading amount 20 µg
Specification Skeletal Muscle
Gel Running Conditions Reduced Denaturing (12% Bis-Tris gel)
Blocking step Li-Cor Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C
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提交于 Dec 20 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Monkey Cell (FRhK-4 cells)
Specification FRhK-4 cells
Fixative Paraformaldehyde
Permeabilization Yes - 0.2%Triton-X100 in PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
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提交于 Nov 07 2012

Vielen Dank für Ihre Anfrage die Herr Hryca uns in den technischen Dienst weitergeleitet hat.

Ich habe geschaut, und wir haben viele möglichen positiven Kontrollen für Sie. Können Sie mir bitte noch sagen, welche Art von p...

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Thank you for your email. Please find all the requested information and discount code below.

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I am very pleased to hear you would like to accept our offer and test ab25361 in IP. This cod...

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Thank you for your reply.

The antibodies differ in the immunogen, tested applications, tested species, purification, concentration, amount and price.

Please see the datasheets for each antibody for this information.
ab25631: ...

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Thank you for contacting us.

The clone was raised against human adherent peripheral blood cells. The epitope it recognizes is unmapped.

Please contact us if you have any other questions.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Huh7)
Specification Huh7
Fixative Paraformaldehyde
Permeabilization Yes - saponin
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 May 21 2012

Thank you for contacting us.
I am sorry to hear about the problems you have encountered.

The bands you are seeing at 25 kDa could be the light chain of endogenous IgG.

As we discussed over the phone, please try the following su...

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