Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24395-31] to LAMP1 - BSA and Azide free
  • Suitable for: WB, ICC, IHC-P, Flow Cyt
  • Reacts with: Human

Overview

  • Product name

    Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free
    See all LAMP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR24395-31] to LAMP1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC, IHC-P, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1; MCF7 whole cell lysate; Human kidney, spleen, colon cancer tissue lysate; HeLa; HepG2 whole cell lysate. IHC-P: Human tonsil, pancreas, liver and lung cancer tissues. ICC: WT HAP1 cells; HeLa cells. Flow Cyt: WT HAP1 cells, HeLa cells.

  • General notes

    ab278046 is the carrier-free version of ab278043. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab278046 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Images

  • Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    All lanes : Anti-LAMP1 antibody [EPR24395-31] (ab278043) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 45 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Blocking and Diluting buffer: 5% NFDM/TBST

    LAMP1 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 27061067, 15111122).

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human liver. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunocytochemistry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling LAMP1 with ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (Green). Confocal image showing cytoplasmic staining in HeLa. ab11267 Anti-LAMP1 mouse monoclonal antibody was used to counterstain at 1/500 (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Flow Cytometry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell, Right)/ LAMP1 knockout HAP1 cells (Left) cells labelling LAMP1 with ab278043 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    All lanes : Anti-LAMP1 antibody [EPR24395-31] (ab278043) at 1/1000 dilution

    Lane 1 : Human kidney tissue lysate
    Lane 2 : Human spleen tissue lysate
    Lane 3 : Human colon cancer tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody(HRP))

    Predicted band size: 45 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Blocking and Diluting buffer: 5% NFDM/TBST

    LAMP1 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 27061067, 15111122).

    Exposure time: 3 minutes.

  • Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Western blot - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    All lanes : Anti-LAMP1 antibody [EPR24395-31] (ab278043) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate
    Lane 2 : LAMP1 knockout HAP1 whole cell lysate
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : Human kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at 1/10000 dilution (Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD))

    Predicted band size: 45 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Blocking buffer and Diluting buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1-4: Merged signal (red and green). Green - ab278043 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab278043 Anti-LAMP1 antibody [EPR24395-31A] was shown to specifically react with LAMP1 in wild-type HAP1 cells. Loss of signal was observed when knockout cell line was used. Wild-type and LAMP1 knockout samples were subjected to SDS-PAGE. ab278043 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human tonsil labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human tonsil. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution  followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human lung cancer. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling LAMP1 with ab278043 at 1/5000 (0.109 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Cytoplasmic staining in human pancreas. The section was incubated with ab278043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Immunocytochemistry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation. 

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LAMP1 KO HAP1 a cells labelling LAMP1 with ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in parental HAP1 cell line and no staining in LAMP1 KO HAP1 cell line. ab11267 Anti-LAMP1 mouse monoclonal antibody was used to counterstain at 1/200 (Red). Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)
    Flow Cytometry - Anti-LAMP1 antibody [EPR24395-31] - BSA and Azide free (ab278046)

    This data was developed using ab278043, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labelling LAMP1 with ab278043 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

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