Overview

  • Product name
    Anti-beta Galactosidase antibody
    See all beta Galactosidase primary antibodies
  • Description
    Chicken polyclonal to beta Galactosidase
  • Tested applications
    Suitable for: ELISA, IHC-FrFl, ICC, IHC-FoFr, Flow Cyt, ICC/IF, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Escherichia coli
  • Immunogen

    Full length native protein (purified). The immunogen was purified beta-galactosidase from Escherichia coli.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    0.01M PBS pH7.4, 15mM Na azide
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antibodies were solid phase absorbed then immunoaffinity purified using purified beta-galactosidase immobilized on a solid phase.
  • Clonality
    Polyclonal
  • Isotype
    IgY
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9361 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
ELISA Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-FoFr 1/1000 - 1/2000.
Flow Cyt Use at an assay dependent concentration.

ab37382 - Chicken polyclonal IgY, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 0.5 µg/ml. Predicted molecular weight: 116 kDa.
IHC-P 1/200 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Relevance
    Beta galactosidase is coded by a gene (lac z) in the lac operon of Escherichia coli. It is a metalloenzyme that splits lactose into glucose and galactose. It hydrolyzes terminal, non-reducing beta-D-galactose residues in beta-D-galactosides. Activation by cations seems to be substrate dependent. K+, Na+, NH4+, Rb+, Cs+ and Mn++ all activate enzyme activity based upon the substrate used.
  • Cellular localization
    Cytoplasmic
  • Database links
    • Alternative names
      • Beta D galactosidase antibody
      • Beta gal antibody
      • Beta galactosidase antibody
      • ECK0341 antibody
      • JW0335 antibody
      • Lactase antibody
      • lacZ antibody
      see all

    Images

    • P0-adult mice were euthanized and perfused with 4% paraformaldehyde in PBS (PF). Their spinal cords were then post-fixed for 30–60 mins in 4% PF at 4°C (P0) or at room temperature (adult). Spinal cords were rinsed and cryoprotected in 20% sucrose in PBS (4°C) prior to embedding in OCT (Tissue-Tek). Immunostaining of frozen spinal sections was performed by incubating 20 µm thick sections with primary antibodies, which were then detected using species-specific secondary antibodies conjugated with Cy2, Cy3 and Cy5 or FITC. ab9361 was used at 1:1000.

    • ab9361 staining beta Galactosidase in mouse e13 stomach and liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with Davidson's fixative, permeabilized with 0.5% Triton-X 100 and blocked with 10% serum for 30 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/500 in TBST + 10% goat serum) for 16 hours at 4°C. A Biotin-conjugated goat anti-chicken IgY polyclonal (1/500) was used as the secondary antibody.

      See Abreview

    • ab9361 staining beta Galactosidase in fruit fly central nervous system glia cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde, permeabilised in 0.1% Triton. Samples were then incubated with primary antibody at 1/2000 for 12 hours at 4°C. The secondary antibody used was a donkey anti-chicken monoclonal conjugated to DyLight® 649 (pink) used at a 1/400 dilution. Nuclei stained with a pan nuclear marker (green).

      See Abreview

    • Ab9361 staining Beta galactosidase in Mouse thyroid tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed 10% buffered formalin, cut into 3-4 micron slices, blocked with 10% normal goat serum and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with the following primary antibodies; anti-BrdU, anti-Sca1 (ab109211, 1∶600), anti-NKX2-1 and anti-β-gal (ab9361, 1∶1000). Sections were incubated with the first primary antibody (anti-Sca1) for 1 hour at room temperature. After washing with PBS, sections were incubated with the first secondary antibody (Alexa Fluor 555 goat anti-rabbit IgG) and washed with PBS. Sections were then incubated with normal serum (5% rabbit serum) from the same host species as the first primary antibodies for 1 hour at room temperature and washed with PBS. Sections were further incubated with an excess of unconjugated Fab antibody derived from the same host species as the primary antibody for 1 hour at room temperature and washed with PBS. The sections were finally incubated with the mixed second primary antibodies (anti-BrdU, anti-β-gal, anti-NKX2-1) overnight at 4°C, washed with PBS, and were incubated with the second secondary antibody (Alexa Fluor 488 goat anti-rat IgG, Dylight 650 goat anti-chicken IgY, Dylight 594 goat anti-rabbit IgG) for 1 hour at room temperature and washed with PBS. DAPI dye was used to stain the nuclei of cells.

    • ab9361 staining Beta Galactosidase in mouse retinal tissue sections by IHC-Fr (Frozen sections). Tissue samples were fixed with formaldehyde, permeabilized with 0.2% triton-X and blocked with 5% serum for 1 hour at 23°C. The sample was incubated with primary antibody (1/1500 in PBS, 2% serum, 0.2% Triton-X) at 4°C for 16 hours. An Alexa Fluor®488-conjugated Goat monoclonal to chicken IgY (1/200) was used as secondary antibody.

      See Abreview

    • ab9361 at 1/250 staining human HEK293T cells by ICC/IF. The cell line was transfected with a b-gal expressing plasmid, and x-gal staining was performed on adjacent wells. The cells were paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 16 hours. A Texas Red conjugated donkey anti-chicken antibody was used as the secondary.

      See Abreview

    References

    This product has been referenced in:

    See all 259 Publications for this product

    Customer reviews and Q&As

    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Username

    Abcam user community

    Verified customer

    Submitted Jan 12 2016

    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 (E12.5 Embryo)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris EGTA, pH=9, pressure cooker
    Permeabilization
    Yes - Saponi
    Specification
    E12.5 Embryo
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Feb 16 2016

    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Username

    Abcam user community

    Verified customer

    Submitted Jan 14 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Fruit fly (Drosophila melanogaster) Cell (Larval brain)
    Permeabilization
    Yes - TBS-T
    Specification
    Larval brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Aug 20 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Intestine)
    Permeabilization
    Yes - .01 SDS
    Specification
    Intestine
    Blocking step
    Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 22 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Main Olfactory Epithelium (MOE))
    Permeabilization
    Yes - TritonX 0.01%
    Specification
    Main Olfactory Epithelium (MOE)
    Blocking step
    Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Jennifer Cisson

    Verified customer

    Submitted May 11 2015

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    None
    Sample
    Mouse Tissue sections (cultured acute spinal cord slice from Roza26-LacZ)
    Specification
    cultured acute spinal cord slice from Roza26-LacZ
    Permeabilization
    Yes - Triton X-100
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 19 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citric
    Sample
    Mouse Tissue sections (e13 stomach and liver)
    Specification
    e13 stomach and liver
    Permeabilization
    Yes - 0.5 % Triton-X 100
    Fixative
    Davidson's fixative
    Username

    Dr. Laurel Baglia

    Verified customer

    Submitted Mar 24 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step
    None
    Sample
    Mouse Tissue sections (Mouse, whole brain sections, striatum)
    Specification
    Mouse, whole brain sections, striatum
    Permeabilization
    Yes - Tween-20
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 08 2013


    ab9361 was raised against E. coli beta galactosidase, which only has 11% homology with mouse beta galactosidase. In general, we like to see greater than 85% homology before we expect an antibody to cross react with an untested species. Thus, ab936...

    Read More

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