Recombinant
RabMAb

Recombinant Anti-Lysozyme antibody [EPR2994(2)] (ab108508)

Rabbit recombinant monoclonal Lysozyme antibody [EPR2994(2)]. Validated in WB, IHC, ICC/IF and tested in Mouse, Human. Cited in 27 publication(s). Independently reviewed in 2 review(s).

Overview

  • Product name
    Anti-Lysozyme antibody [EPR2994(2)]
    See all Lysozyme primary antibodies
  • Description
    Rabbit monoclonal [EPR2994(2)] to Lysozyme
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human Lysozyme aa 50-150. The exact sequence is proprietary.

  • Positive control
    • WB: RAW 264.7 and HL-60 whole cell lysate; Human spleen tissue lysate; Natural human Lysozyme protein. IHC-P: Human tonsil, spleen, lung, kidney, brain, breast and heart tissues; Mouse spleen and small intestine tissues. ICC/IF: THP-1 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 ab108508 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/IF 1/250.

 For unpurified use at 1/500 - 1/1000.

WB 1/10000 - 1/50000. Predicted molecular weight: 17 kDa.
IHC-P 1/1000 - 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.

See IHC antigen retrieval protocols.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Lysozymes have primarily a bacteriolytic function; those in tissues and body fluids are associated with the monocyte-macrophage system and enhance the activity of immunoagents.
    • Involvement in disease
      Amyloidosis 8
    • Sequence similarities
      Belongs to the glycosyl hydrolase 22 family.
    • Cellular localization
      Secreted.
    • Information by UniProt
    • Database links
    • Alternative names
      • 1 4 beta N acetylmuramidase C antibody
      • 1 antibody
      • 4-beta-N-acetylmuramidase C antibody
      • EC 3.2.1.17 antibody
      • LYSC_HUMAN antibody
      • Lysosyme antibody
      • Lysozyme (renal amyloidosis) antibody
      • Lysozyme C antibody
      • Lysozyme C precursor antibody
      • LYZ antibody
      • LZM antibody
      • Renal amyloidosis antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded human lung tissue sections labeling lysozyme with purified ab108508 at a dilution of 1/1500 (0.6 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody.Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using EDTA Buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

    • Anti-Lysozyme antibody [EPR2994(2)] (ab108508) at 1/10000 dilution (purified) + RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 17 kDa
      Observed band size: 16 kDa
      why is the actual band size different from the predicted?



      Blocking/Diluting buffer 5% NFDM/TBST
    • Immunocytochemistry/Immunofluorescence analysis of THP-1 (Human monocytic leukemia) cells labeling lysozyme with purified ab108508 at 1/250. Cells were fixed with 100% methanol. ab150077, Alexa Fluor®488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 g/mL) - Microtubule Marker (Alexa Fluor®594) at 1/200. DAPI (blue) was used as a nuclear counterstain. Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue sections labeling lysozyme with purified ab108508 at a dilution of 1/1500 (0.6 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using EDTA Buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

    • All lanes : Anti-Lysozyme antibody [EPR2994(2)] (ab108508) at 1/10000 dilution (unpurified)

      Lane 1 : HL60 (human promyelocytic leukemia cell line) cell lysate
      Lane 2 : Human spleen lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 17 kDa

    • Paraffin-embedded mouse small intestine tissue stained for Lysozyme using ab108508 in immunohistochemical analysis.

      See Abreview

    • Anti-Lysozyme antibody [EPR2994(2)] (ab108508) at 1/50000 dilution (purified) + HL-60 (Human promyelocytic leukemia cell line) whole cell lysate at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 17 kDa
      Observed band size: 16 kDa why is the actual band size different from the predicted?



      Blocking/Diluting buffer 5% NFDM/TBST
    • Unpurified ab108508, at 1/1000 dilution, staining Lysozyme in Human tonsil by Immunohistochemistry, Paraffin-embedded tissue.

    • Anti-Lysozyme antibody [EPR2994(2)] (ab108508) at 1 µg/ml (unpurified) + Native human Lysozyme protein (ab91125) at 0.1 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 17 kDa


      Exposure time: 4 minutes
    • Unpurified ab108508, at 1/1000 dilution, staining Lysozyme in Human spleen by Immunohistochemistry, Paraffin-embedded tissue.

    • Unpurified ab108508, at 1/1000 dilution, staining Lysozyme in Human kidney by Immunohistochemistry, Paraffin-embedded tissue.

    • Unpurified ab108508 showing negative staining in Normal brain tissue.

    • Unpurified ab108508 showing negative staining in Normal breast tissue.

    • Unpurified ab108508 showing negative staining in Normal heart tissue.

    References

    This product has been referenced in:
    • Kriesel JD  et al. Spectrum of Microbial Sequences and a Bacterial Cell Wall Antigen in Primary Demyelination Brain Specimens Obtained from Living Patients. Sci Rep 9:1387 (2019). Read more (PubMed: 30718694) »
    • Zhang X  et al. Interleukin-22 regulates the homeostasis of the intestinal epithelium during inflammation. Int J Mol Med 43:1657-1668 (2019). Read more (PubMed: 30816423) »
    See all 33 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (lungs)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA
    Permeabilization
    No
    Specification
    lungs
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 08 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
    Permeabilization
    No
    Specification
    Spleen
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 31 2018

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (small intestine)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate
    Permeabilization
    No
    Specification
    small intestine
    Blocking step
    AB block Dako X0590 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    10% NBF

    Mr. Thorsten Cramer

    Verified customer

    Submitted Aug 18 2016

    Answer

    This antibody would not distinguish between the mouse M or P lysozyme isoforms as the immunogen sequence in conserved in both.

    Read More

    Answer

    Thank you for providing that information.

    I have had a look at the two rabbit monoclonal against Lysozyme (ab91653 and ab108508). Both of these antibodies should be able to detect all three forms of the protein (the wild type as well as your two mutants). The immunogen used to raise the ab108508 antibody has been taken from a region N-terminal of your mutations, whilst the immunogen used to raise the antibody ab91653 is from the C-terminal domain. Neither immunogen overlaps any of the mutation sites.

    I hope this information has beenof help. If you have any further questions please do not hesitate to ask.

    Read More

    Question
    Answer


    Thank you for contacting us yesterday.

    I have received further information from the lab regarding the IHC protocol for ab108508. I can
    confirm that 0.01M Sodium Citrate Buffer, pH 6.0 was used for antigen retrieval. Here is a copy
    of the protocol:

    Immunohistochemistry Protocol for Paraffin-embedded Tissues

    1. Solutions and reagents
    1.1. Xylene
    1.2. Ethanol, anhydrous denatured, histological grade (100%, 95%, 70%, 50%)
    1.3. Washing buffer/TBST: 1X TBS/0.1% Tween-20, pH to 7.6.
    1.4. Distilled water (dH2O)
    1.5. Antigen Retrieval Solution: 0.01M Sodium Citrate Buffer, pH 6.0
    To prepare Antigen Retrieval stock solutions:
    10X Stock: Dissolve 29.4 g sodium citrate trisodium salt dehydrate (C 6H 5Na 3O 7 2H2O in 1 liter of dH2O. Add 5mL Tween-20.
    1X Working Solution: Mix 200mL 10X stock with 1800mL dH2O; pH to 6.0
    1.6. 3% Hydrogen Peroxide
    1.7. Blocking Buffer: 10% serum in PBS (serum origin depends on the host of the secondary antibody)
    1.8. Primary Antibody Diluent: 5% serum in PBS (serum origin depends on the host of the
    Secondary antibody)
    1.9. Hematoxylin
    1.10. Permanent Mounting Medium

    2. Protocol
    2.1. Deparaffinization/Rehydration
    2.1.1. Heat slides in an oven at 65 °C for 1 hour.
    2.1.2. De-paraffinize/hydrate using the following series of washes: two Xylene washes (3 min each), followed by two 100% ethanol rinses (3 min each), followed by 95% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, followed by TBST wash for 3 min on a shaker.

    2.2. Antigen Retrieval
    This is recommended Heat Induced Epitope Retrieval (HIER) using Decloaking
    Chamber/Pressure Cooker. Hot water bath or Microwave with temperature sensor can be also used
    (protocol would vary depending on the method used).
    2.2.1. Add 500 ml of dH 2O to Decloaker/Pressure Cooker.
    2.2.2. Immerse slides into staining dish containing Antigen Retrieval Solution. Place staining dish into decloaking chamber.
    2.2.3. Program to run for 30 seconds at 125° C, followed by 10 seconds at 90° C.
    2.2.4. Let it cool down to room temperature (10 – 20 minutes).
    2.2.5. Removes slides and rinse in TBST.
    2.2.6. Proceed to Staining step.

    2.3. Staining
    2.3.1. Wash slides with TBST for 3 min on a shaker.
    2.3.2. Inactivate endogenous peroxidase by covering tissue with 3% hydrogen peroxide for 5 min.
    2.3.3. Wash slides three times with TBST (3 min each on a shaker).
    2.3.4. Block slides with the blocking solution for 1 hour.
    2.3.5. Dilute primary antibody in primary antibody diluent per recommendation on data sheet.
    2.3.6. Apply primary antibody to each section and incubate overnight in the humidified chamber (4 ºC).
    2.3.7. Wash slides three times with TBST (3 min each on a shaker).
    2.3.8. Apply to each section secondary HRP-conjugated anti-rabbit antibody diluted in the blocking solution per manufacturer’s recommendation; incubate for 30 min at room temperature.
    2.3.9. Wash slides three times with TBST (5 min each on a shaker).
    2.3.10. Add freshly prepared DAB substrate to the sections and incubate until stain develops (generally 1 min).
    2.3.11. Rinse sections with water.
    2.3.12. Counterstain with Hematoxylin (generally 10 seconds).
    2.3.13. Rinse sections with water.
    2.3.14. Dehydrate samples using two washes with 100% Ethanol (3 min each), followed by
    two rinses with Xylene (3 min each).
    2.3.15. Mount coverslips on slides using permanent mounting medium.

    I hope this information helps and that the antibody works well. Please do not hesitate to contact us should you have further problems.

    Read More

    Answer

    Thank you for contacting us. Based on sequence analysis, immunogen of ab91653 does not share any significant sequence similarity with chicken. For the immunogen of ab108508, it shares 67% identity with chicken Lysozyme C, so it should also be human specific. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up