Overview

  • Product name
    Anti-LAMP1 antibody [1D4B]
    See all LAMP1 primary antibodies
  • Description
    Rat monoclonal [1D4B] to LAMP1
  • Host species
    Rat
  • Specificity
    Murine CD107a/LAMP1 (Mr 110-140 kDa with a core protein of Mr ~40 kDa)
  • Tested applications
    Suitable for: IHC-P, ICC/IF, Other, Flow Cyt, IP, IHC-Fr, WB, Immunomicroscopymore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Plasma membrane fraction of mouse embryo NIH3T3 cell line.

  • Positive control
    • IHC-P: human lung FFPE tissue sections

Applications

Our Abpromise guarantee covers the use of ab25245 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
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration. PubMed: 17215451
Other Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

AP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 40 kDa (predicted molecular weight: 45 kDa).
Immunomicroscopy Use at an assay dependent concentration.

Target

  • Function
    Presents carbohydrate ligands to selectins. Also implicated in tumor cell metastasis.
  • Sequence similarities
    Belongs to the LAMP family.
  • Post-translational
    modifications
    O- and N-glycosylated; some of the 18 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
  • Alternative names
    • 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

Images

  • IHC image of LAMP1 staining in human lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab25245, 10µg/ml, for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ab25245 staining LAMP1 in Mouse epithelial cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with methanol/acetone 1:1 and blocked with 1% milk in PNB for 1 hour at 25°C. Samples were incubated with primary antibody (1/500) in PNB for 16 hours at 4°C. An Alexa Fluor® 568-conjugated Donkey polyclonal to rat IgG, dilution 1/500, was used as secondary antibody. Nuclei were stained blue.

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab25245 (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 (ab25245, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ab25245 staining mouse macrophage, hepatoma cell by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 25°C.  The primary antibody was diluted 1/1000 and incubated with the sample for 16 hours at 4°C.  An Alexa Fluor® 594 conjugated goat anti-mouse antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Zbesko JC  et al. Glial scars are permeable to the neurotoxic environment of chronic stroke infarcts. Neurobiol Dis 112:63-78 (2018). Read more (PubMed: 29331263) »
  • Chen D  et al. Chloroquine modulates antitumor immune response by resetting tumor-associated macrophages toward M1 phenotype. Nat Commun 9:873 (2018). Read more (PubMed: 29491374) »
See all 51 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Question
Answer

The predicted calculated MW for LAMP1 is approximately 44.8 kDa (human) and 43.8 kDa (mouse). However, the observed MW for LAMP1 as run on an SDS-PAGE gel can be much higher, and the protein is heavily glycosylated in both species. As such, the predicted observed MW for LAMP1 in both human and mouse is anticipated to be approximately in the range of 110-140 kDa.

Additional information on LAMP1 and its post-translational modifications can be found here:

UniProt Human LAMP1 P11279

UniProt Mouse LAMP1 P11438

Read More

Answer

Unfortunately, we have not performed epitope mapping on product Anti-LAMP1 antibody [1D4B] (ab25245).

However, QC testing has demonstrated that ab25245 recognizes an extracellular epitope on mouse epithelial cells and macrophages.

Similarly, the following alternate anti-LAMP1 Ab product is noted to recognize an extracellular epitope on human activated platelets and U937 cells: Anti-LAMP1 antibody [H4A3] (ab25630)

Read More
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

Abcam user community

Verified customer

Submitted Apr 05 2017

Abcam has not validated the combination of species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Apr 05 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

Abcam user community

Verified customer

Submitted Apr 05 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (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

Abcam user community

Verified customer

Submitted Apr 05 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 24 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain (microgliosis neocortex))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Permeabilization
Yes - Tween20
Specification
brain (microgliosis neocortex)
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 10 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse macrophage)
Permeabilization
No
Specification
mouse macrophage
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Sample
Mouse Cell (Neuronal progenitor cell line)
Specification
Neuronal progenitor cell line
Permeabilization
Yes - PBS 1% BSA 0,1% Saponine
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 08 2014

1-10 of 15 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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