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Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)

Reviews (19)Q&A (8)References (175)
Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Flow Cytometry (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
  • Flow Cytometry (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)

Key features and details

  • Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker
  • Suitable for: ICC, ICC/IF, IHC-Fr, WB, IHC-FoFr, Flow Cyt (Intra), IHC-P
  • Reacts with: Human, Rhesus monkey, African green monkey
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Biotin PE PE/Cy5®

Overview

  • Product name

    Anti-LAMP2 antibody [H4B4] - Lysosome Marker
    See all LAMP2 primary antibodies

  • Description

    Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC, ICC/IF, IHC-Fr, WB, IHC-FoFr, Flow Cyt (Intra), IHC-Pmore details

  • Species reactivity

    Reacts with: Human, Rhesus monkey, African green monkey
    Does not react with: Mouse, Rat

  • Immunogen

    Tissue, cells or virus corresponding to Human LAMP2. Human adherent peripheral blood cells

  • Positive control

    • Flow Cyt (Intra): THP1 cells.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC (1)
Use at an assay dependent concentration.
ICC/IF (11)
Use at an assay dependent concentration.
IHC-Fr (1)
Use at an assay dependent concentration.
WB (5)
Use at an assay dependent concentration.

In our hands milk blocking is gives superior results to BSA blocking for this product.
IHC-FoFr
Use at an assay dependent concentration.

(PMID 19837699).
Flow Cyt (Intra)
Use 0.5-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P (1)
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
ICC
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IHC-Fr
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-FoFr
Use at an assay dependent concentration.

(PMID 19837699).
Flow Cyt (Intra)
Use 0.5-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    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.

  • Tissue specificity

    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.

  • Involvement in disease

    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.

  • Sequence similarities

    Belongs to the LAMP family.

  • Post-translational
    modifications

    O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.

  • Cellular localization

    Cell membrane. Endosome membrane. Lysosome membrane. This protein shuttles between lysosomes, endosomes, and the plasma membrane.
  • Information by UniProt

  • Database links

    • Form

      Alternative splicing produces 3 isoforms.

    • Alternative names

      • CD107 antigen like family member B antibody
      • 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
      • LGP 96 antibody
      • LGP110 antibody
      • LGP96 antibody
      • Lysosomal associated membrane protein 2 antibody
      • Lysosome associated membrane glycoprotein 2 antibody
      • Lysosome associated membrane protein 2 antibody
      • Lysosome-associated membrane glycoprotein 2 antibody
      • Lysosome-associated membrane protein 2 antibody
      • MAC3 antibody
      see all

    Images

    • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      All lanes : Anti-LAMP2 antibody [H4B4] - Lysosome Marker (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
      All lanes : 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      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.
    • Flow Cytometry (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Flow Cytometry (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)

      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.

       

    • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)This image is of an Abreciew submitted by Daniel Rodriguez.
      All lanes : Anti-LAMP2 antibody [H4B4] - Lysosome Marker (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
      All lanes : HRP conjugated Goat anti-Mouse IgG at 1/2000 dilution

      Developed using the ECL technique.

      Exposure time: 30 seconds


      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

    • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)This image was kindly supplied by Daniel Rodriguez by Abreview
      Anti-LAMP2 antibody [H4B4] - Lysosome Marker (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


      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

    • Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)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 (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
      Flow Cytometry (Intracellular) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)

      Intracellular 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. 

       

       

    References (175)

    Publishing research using ab25631? Please let us know so that we can cite the reference in this datasheet.

    ab25631 has been referenced in 175 publications.

    • Palomba NP  et al. Common and Rare Variants in TMEM175 Gene Concur to the Pathogenesis of Parkinson's Disease in Italian Patients. Mol Neurobiol 60:2150-2173 (2023). PubMed: 36609826
    • Krishnan B  et al. Active RB causes visible changes in nuclear organization. J Cell Biol 221:N/A (2022). PubMed: 35019938
    • Wang S  et al. Activation of Transcription Factor EB Alleviates Tubular Epithelial Cell Injury via Restoring Lysosomal Homeostasis in Diabetic Nephropathy. Oxid Med Cell Longev 2022:2812493 (2022). PubMed: 35082964
    • Kumar R  et al. Combinatorial Polycation Synthesis and Causal Machine Learning Reveal Divergent Polymer Design Rules for Effective pDNA and Ribonucleoprotein Delivery. JACS Au 2:428-442 (2022). PubMed: 35252992
    • Fukushima S  et al. Recruitment of LC3 by Campylobacter jejuni to Bacterial Invasion Site on Host Cells via the Rac1-Mediated Signaling Pathway. Front Cell Infect Microbiol 12:829682 (2022). PubMed: 35310852
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-8 of 8 Q&A

    Question

    Sehr geehrter Herr Hryca,
    vielen lieben Dank für das Angebot. Wir würden gerne die Maus-Antikörper
    bestellen und dabei ist uns aufgefallen, dass wir keine Möglichkeit einer
    Positivkontrolle haben. Können Sie uns da vielleicht weiterhelfen?
    Mit freundlichen Grüßen aus Düsseldorf

    Read More
    Answer

    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 positive Kontrolle Sie benötigen? Ist es ein Lysat für Western blot oder ein Gewebeschnitt für Immunohistochemie? Haben Sie auch eine Präferenz für eine Spezies (Maus, human)?

    Hier mal eine Aufstellung von möglichen positiven Kontrollen. Ich freue mich wieder von Ihnen zu hören, um Ihnen genaure Vorschläge machen zu können.




    cell lines Slide
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=2105 Rabbit polyclonal to IL1 beta LPS treated RAW 264.7 cells, Alveolar macrophages (MHS cell line) liver, diseased, inflammed tissue, monocytes, macrophages
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6313 Mouse monoclonal [CTD-19] to Cathepsin D SKBR3, MCF7 Human liver tissue.
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6535 Mouse monoclonal [BRM-22] to Hsp70 Hela, 293T,MCF7, Jurkat, HepG2, A431, HCT116, PC3, ... liver, brain, testes, heart, skeletal muscle
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=8219 Mouse monoclonal [MEM-259] to CD63 HT1080, HepG2, OVCAR-3, HL60 and A375 hemapoietic ells, tissue macrophages, platelet, human melanoma tissue, monocytes, macrophages, endothelial cells, SM cells, brain white matter, peripheral nerves
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=25631 Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker HepG2, 3T3, HeLa, RAW264.7, ... placenta, lung, liver
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=50533 Mouse monoclonal [Rab7-117] to RAB7 HepG2, B16, SK-MEL-28, 3T3, Human brain whole tissue, mouse neuroblastoma
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=68340 Mouse monoclonal to Rab9 Hela, A172, 3T3, 293, PC-12. COS-7, THP-1 tonsil, spleen, prostate cancer, stomach tissu
    http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=72907 Rabbit polyclonal to Rab5b Hela, Jurkat, HEK293, HepG2 Human Placenta Tissue, Human Brain Tissue.

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    Question

    Thank you for your response.

    I am glad to take your advice to order ab25361 (Anti-LAMP2 antibody [H4B4] - Lysosome Marker) from you. And I ‘d like to take part in the testing program. So please mail me the discount code.

    What’s more, I wander when I should use the code, at the time I order ab25361 or the next time I order other things from you?

    Read More
    Answer

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

    DISCOUNT CODE: xxx
    Expiration date: xxx

    I am very pleased to hear you would like to accept our offer and test ab25361 in IP. This code will give you: 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for IPand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
    For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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    Question

    I am so glad to receive a quick response from you concerning about the LAMP-2 antibody. I want to order some mouse antibody, which you listed as ab25631, ab119124 and ab112019. Could you tell me the differences among the three antibody? I really what to get the detailed information about them, and choose the best one for my Co-IP assay.

    What’s more I ‘d like to take part in the testing discount program, so please send me the code for it.

    Read More
    Answer

    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: https://www.abcam.com/lamp2-antibody-h4b4-lysosome-marker-ab25631.html
    ab119124: https://www.abcam.com/lamp2-antibody-h4b4-azide-free-ab119124.html
    ab112019: https://www.abcam.com/lamp2-antibody-h4b4-ab112019.html

    My recommendationwould be touse ab25631 for the IP testing as it has been used in many different applications as well. It has been referenced in publications as well as customers have submitted their feedback in form of Abreviews (https://www.abcam.com/LAMP2-antibody-H4B4-Lysosome-Marker-ab25631-reviews.html).

    I'd be happyto send you a testing discount code. Please let me know for which antibody you would like to receive the code.

    I look forwad to hear back from you.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Question

    I am using the anti body ab25631-110 anti LAMP2 for WB and immunofluorescence. I wold like to know what part of LAMP2 this anti body recognize, the C terminal cytosolic portion or the N terminal lumenal portion?

    Read More
    Answer

    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.

    Read More

    Question

    ab25630: WB with rat liver
    sees band at 25 kDa
    transfer is OK, detects other proteins in the blot
    Ab: 1/1000 1-2 h RT
    block: overnight 4 degree
    secondary: OK
    antibody from other company to same protein gives same results
    suggestions: WB conditions might not yet be optimal, try blocking for only 1-2 h RT, antibody incubation: overnight 4 degree
    ab25631: rat is unsuitable
    ab2773: WB with rat liver, sees band at 25 kDa
    suggestions: WB conditions might not yet be optimal, try blocking for only 1-2 h RT, antibody incubation: overnight 4 degree

    Read More
    Answer

    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 suggestions as it seems the WB conditions for these 2 antibodies (ab25630 and ab2773) are not yet optimal:

    1) try blocking for only 1-2 h RT
    2) try the primary antibody incubation for overnight 4 degree

    As for ab25631, this has been shown to not react with rat samples and would probably not give satisfactory results.However, if you have access to a human sample, you could try the antibody on that.

    Please let me know if the tips were helping to improve your results.

    I hope this information is helpful to you, and I look forward to hear back.

    Read More

    Question

    Hi, The power cut was not long than 24 hours and we cannot assure if the products were or not at room temperature when the electricity switched again (they were kept in the fridge but it was not working as a consequence of the power cut). We need your evaluation about the state of the products, because if they are damaged we will claim it cost to the insurance company and we will process with its reposition. Thanks in advance for your help and collaboration Kind regards,    

    Read More
    Answer

    Thank you for contacting us. My colleague "Ariana" has already sent her reply so I am closing this case. The antibodies are fine for 1 week at room temperature however in your case if the products fail to perform I would suggest getting refund from insurance company. You can get information of product price from our website at  https://www.abcam.com/ I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.    

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    Question

    Dear Sir/Madame, Recently IRB Barcelona suffered a power cut and it caused that refrigerators and freezers located in our plant were out of operating. In order to proceed with an official claim for the possible costs that could arise from damaged products as a result of their inadequate storage, we will appreciate your evaluation about the items or products that should be rejected and destroyed because their proper running could not being guaranteed. Please find below the items/products affected Supplier Ref. ITEM Description Abcam ab25631 Ms mAb to LAMP2[H4B4] Abcam ab47517 mtTFA antibody (ab47517 Abcam ab56889 Anticòs monoclonal Mfn2     Thanks in advance for your help and cooperation.   Kind regards.      

    Read More
    Answer

    Thank you for contacting us. In order to help you with your question could you provide the following - Order number of purchase - for how long the power cut happened? -Were these products at room temperature when the electricity switched on again? What do you really want us to do? Do you need quote for these products please explain? I look forward to hearing from you soon.

    Read More

    Question

    Does antibody H4B4 recognize LAMP-2B?

    Read More
    Answer

    Thank you for contacting us. In humans, the three splice variants (LAMP-2A, LAMP-2B and LAMP-2C) all have identical extracellular domains. It is also known that LAMP-2A is distributed rather ubiquitously, while LAMP-2B is expressed predominantly in striated muscle and brain. Clone H4B4 has been shown to stain for LAMP-2 in skeletal muscle. This information leads us to believe that this antibody will react with LAMP-2B, though we have not verified this through in-house testing. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    For licensing inquiries, please contact partnerships@abcam.com