Recombinant
RabMAb

Recombinant Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

Reviews (3)Q&A (1)References (13)

Overview

  • Product name

    Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker
    See all LAMP2A primary antibodies

  • Description

    Rabbit monoclonal [EPR4207(2)] to LAMP2A - Lysosome Marker

  • Host species

    Rabbit

  • Specificity

    ab125068 only recognizes LAMP2a.

  • Tested applications

    Suitable for: Flow Cyt, WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LAMP2A aa 400 to the C-terminus.
    Database link: P13473-1

  • Positive control

    • WB: HeLa, Jurkat, ECV-304, JAR, NIH/3T3, RAW 264.7, PC-12 and C6 whole cell lysates. Mouse kidney lysates and Rat liver and kidney lysates. IHC-P: Human placenta and liver tissue. Mouse and rat liver tissue. IP: HeLa and RAW 264.7 whole cell lysates. Flow Cyt: HeLa cells.

  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab125068 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
Flow Cyt 1/1000.
WB 1/2000. Detects a band of approximately 120 kDa (predicted molecular weight: 45 kDa).
IP 1/60 - 1/100.
IHC-P 1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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

    Danon disease

  • 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

    see all

  • Form

    Alternative splicing produces 3 isoforms.

  • Alternative names

    • CD107 antigen-like family member B antibody
    • CD107b antibody
    • LAMP 2 antibody
    • Lamp 2a antibody
    • LAMP-2 antibody
    • LAMP2 antibody
    • LAMP2_HUMAN antibody
    • Lysosome-associated membrane glycoprotein 2 antibody
    • Lysosome-associated membrane protein 2 antibody
    see all

Images

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (purified)

    Lane 1 : Mouse kidney tissue lysate
    Lane 2 : Rat kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Difference in MW may be caused by different degree of glycosylation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Flow Cytometry - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

  • Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/10000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : ECV-304 cell lysate
    Lane 3 : JAR cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    May be seen at ~50 kDa representing the unglycosylated isoforms of LAMP2 and ~120 kDa representing the glycosylated form.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

  • Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

     

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/5000 dilution (unpurified) + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunoprecipitation - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/5000 dilution (unpurified)

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : ECV-304 (Human urinary bladder cancer cell line) whole cell lysate
    Lane 3 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (unpurified)

    Lane 1 : Mouse kidney
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    Western blot - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
    All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (unpurified)

    Lane 1 : Rat liver
    Lane 2 : Rat kidney
    Lane 3 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa


    Exposure time: 3 minutes


    Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:

  • Zhu ZC  et al. Targeting KPNB1 overcomes TRAIL resistance by regulating DR5, Mcl-1 and FLIP in glioblastoma cells. Cell Death Dis 10:118 (2019). Read more (PubMed: 30742128) »
  • Liu DX  et al. Exosomes derived from HBV-associated liver cancer promote chemoresistance by upregulating chaperone-mediated autophagy. Oncol Lett 17:323-331 (2019). Read more (PubMed: 30655770) »
See all 13 Publications for this product

Publishing research using ab125068? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker

 Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing
Loading amount
5 µg
Specification
HEK293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Nov 28 2016

Immunohistochemistry (Frozen sections) abreview for Anti-LAMP2a antibody [EPR4207(2)] - Lysosome Marker

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Rat Tissue sections (brain)
Specification
brain
Permeabilization
Yes - triton-100
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 25 2014

Western blot abreview for Anti-LAMP2a antibody [EPR4207(2)] - Lysosome Marker

 Below Average
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Human Cell lysate - whole cell (BA9 - Cortical area)
Specification
BA9 - Cortical area
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Dr. Martin Broadstock

Verified customer

Submitted Aug 19 2013

Question

Would it be possible to get the concentrations of the current lots of the following Abs?

3768-1

3660-1

Read More
Answer

The IgG concentrations of the current lots are:

3768-1 (ab109534, lot GR68319) - 0.032 mg/ml

3660-1 (ab125068, lot GR75050) - 1.554 mg/ml

Read More

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