Anti-LAMP1 antibody - Lysosome Marker (ab24170)

Overview

  • Product nameAnti-LAMP1 antibody - Lysosome MarkerSee all LAMP1 primary antibodies ...
  • Description
    Rabbit polyclonal to LAMP1 - Lysosome Marker
  • Tested applicationsIP, IHC-P, ICC, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Hamster, Cat, Dog, Human, Xenopus laevis, Zebrafish
    Predicted to work with: Cow
  • Immunogen

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

    (Peptide available as ab25744.)

  • Positive control
    • This antibody gave a positive result in the following whole cell lysates: Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line) MCF7 (Human breast adenocarcinoma cell line)

Properties

Applications

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.

Application Abreviews Notes
IP Use at an assay dependent dilution. PubMed: 21152024
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC Use at an assay dependent dilution.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).Can be blocked with Human LAMP1 peptide (ab25744).
IHC-Fr Use at an assay dependent concentration.

Target

  • FunctionPresents carbohydrate ligands to selectins. Also implicated in tumor cell metastasis.
  • Sequence similaritiesBelongs to the LAMP family.
  • Post-translational
    modifications
    O- and N-glycosylated; some of the 18 N-linked glycans are polylactosaminoglycans.
  • Cellular localizationCell 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

Anti-LAMP1 antibody - Lysosome Marker images

  • 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 AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green) and AlexaFluor®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.

     

    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.

  • 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.
  • 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

  • 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

  • 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

  • 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.

  • 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

References for Anti-LAMP1 antibody - Lysosome Marker (ab24170)

This product has been referenced in:
  • Sun Y  et al. Autophagy benefits the replication of Newcastle disease virus in chicken cells and tissues. J Virol 88:525-37 (2014). Read more (PubMed: 24173218) »
  • Allen SJ  et al. Binding of HSV-1 glycoprotein K (gK) to signal peptide peptidase (SPP) is required for virus infectivity. PLoS One 9:e85360 (2014). ICC/IF . Read more (PubMed: 24465545) »

See all 41 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 22°C
Sample Monkey Cell (COS7)
Specification COS7
Permeabilization Yes - 0.2%Triton
Fixative Formaldehyde
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Submitted May 20 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (h9c2)
Specification h9c2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Jan 02 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (mouse heart)
Specification mouse heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Jan 02 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Jan 02 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 21°C
Sample Chicken Cultured Cells (Retinal Ganglion Cell culture)
Specification Retinal Ganglion Cell culture
Permeabilization Yes - PBS + 0.1% Tween20
Fixative Paraformaldehyde
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Submitted Nov 04 2013

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Mouse Tissue sections (uterus)
Specification uterus
Permeabilization No
Fixative Formaldehyde
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Dr. ananoumous ananoumous

Verified customer

Submitted Sep 03 2013

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (4-12)
Sample Mouse Tissue lysate - whole (uterus, placenta)
Specification uterus, placenta
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Dr. ananoumous ananoumous

Verified customer

Submitted Aug 07 2013

I have attached a paper by Michael Neumann and Detlef Gabel outlining ways in which to reduce fluorescent background. They recommend irradiating tissue before treatment with fluorescent probes. This seems like a simple and possibly quite effective way ...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Sample Human Tissue sections (BA9 cortical sections)
Specification BA9 cortical sections
Permeabilization Yes - 0.05% Tween-20
Fixative Paraformaldehyde
Username

Dr. Martin Broadstock

Verified customer

Submitted Jun 05 2013

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

Dr. Martin Broadstock

Verified customer

Submitted Jun 03 2013

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