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Products:Tags & Cell Markers >> Fusion / Marker Proteins >> Beta Galactosidase
Overview
Product name
Anti-beta Galactosidase antibody
See all beta Galactosidase products (18) ...
Description
Rabbit polyclonal to beta Galactosidase
Specificity
This polycolonal antibody reacts with beta-galactosidase from E.coli showing a 464 kDa band (tetramer) under non-reducing conditions of SDS-PAGE and 116 kDa band under reducing conditions. This antibody does not cross-react with mouse beta-galactosidase.
Tested applications
ELISA, WB, IPmore details
Cross reactivity
Reacts with
Escherichia coli
Immunogen
Recombinant full length protein (E. coli).
Positive control
1.0µg of E.coli in a standard sandwich ELISA. Assay by immunoelectrophoresis resulted in a single precipitin arc against purified and partially purified E.coli beta-galactosidase.
General notes
Application Note: This antibody shows inconsistent results between batches in ICC/IF, IF, IHC-P, IHC-Fr and IHC-FoFr. We have received several good Abreviews on these applications, however, we are unable to guarantee positive results. If the antibody is used for IHC, please try fixing with methanol and block with gelatine instead with BSA or NGS. After each antibody incubation, please wash 5x with TBS instead of PBS.We would recommend ab4761 or ab12081 as alternatives. Detects a band of approximately 464 kDa.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Constituents: 1% BSA, 0.15M Sodium chloride, 0.02M Potassium phosphate, pH 7.2
Concentration
Concentration information loading...
Purity
IgG fraction
Purification notes
IgG fraction purified from monospecific antiserum by a multi-step process of delipidation, salt fractionation and ion exchange chromatography and dialysis.
Clonality
Polyclonal
Isotype
IgG
Research areas
Tags & Cell Markers >> Fusion / Marker Proteins >> Beta Galactosidase
Applications
Show applications key
Our Abpromise guarantee covers the use of ab616 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: 1/5000 - 1/25000.(sELISA tested against 1ug E.coli beta galactosidase.)
WB: 1/5000 - 1/25000.Detects a band of approximately 464 kDa.(This antibody shows inconsistent results between batches in ICC/IF, IF, IHC-P, IHC-Fr and IHC-FoFr. We have received several good Abreviews on these applications, however, we are unable to guarantee positive results. If the antibody is used for IHC, please try fixing with methanol and block with gelatine instead with BSA or NGS. After each antibody incubation, please wash 5x with TBS instead of PBS.We would recommend ab4761 or ab12081 as alternatives.)
IP
IP: 1/500 - 1/5000.IP: 1/500 - 1/5000.
Application notes
Is unsuitable for ,ICC/IF,IF,IHC-FoFr,IHC-Fr or IHC-P.
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
Alternative names
- Beta D galactosidase antibody
- Beta gal antibody
- Beta galactosidase antibody
see all
Database links
- Entrez Gene: 945006 Escherichia coli
- SwissProt: P00722 Escherichia coli
Anti-beta Galactosidase antibody images:
Immunocytochemistry/ Immunofluorescence - beta Galactosidase antibody (ab616)
ab616 at 1/400 staining mouse Rosa-Mesenchymal stem cells by ICC/IF.
Mesenchymal stem cells (MSC) were collected from the bone marrow of Rosa mice for b-galactosidase staining. Rosa mice are a line of transgenic mice expressing the LacZ gene controlled by a promoter, which directs expression to essentially all tissues (Rosa 26, Jackson Laboratories). The cells were 2% paraformaldehyde ffixed for 15 minutes and then permealized with Triton-X100 (0.1%) for 10 minutes and then incubated with ab616 overnight at 4°C. The image demonstrates b-galactosidase expressing MSC (green-cytoplasmic localization) and DAPI (blue-nuclear counter stain).
This image is courtesy of an Abreview submitted by Dr Mal Niladri
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - beta Galactosidase antibody (ab616)
ab616 staining beta galactosidase in E15.5 mouse lung tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin embedded tissue sections). Subsequent to deparaffinization the tissue sections were fixed in 4% paraformaldehyde in PBS, hydrated, heated in 10mM citrate buffer (pH 6.0), treated with 1% H2O2 in methanol for 10 min and blocked with 10% of normal serum. The primary antibody was incubated with sample at 4°C overnight.
Image from Xing Yiming et. al., Development 137:825-33 (2010) Fig 1B.
Immunocytochemistry/ Immunofluorescence - beta Galactosidase antibody (ab616)
ab616 staining beta Galactosidase in Human MiaPaCa cells transiently transfected with pCMV-LacZ by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/200 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
This image is courtesy of an anonymous Abreview
Immunohistochemistry (Frozen sections) - beta Galactosidase antibody (ab616)
ab616 staining beta Galactosidase in skin sections of Notch2LacZ knock-in mouse by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. X-gal staining is shown to confirm that beta-gal antibody detects correct signal.
This image is courtesy of an anonymous Abreview
Western blot - beta Galactosidase antibody (ab616)
Anti-beta Galactosidase antibody (ab616) at 1/5000 dilution + whole cell lysate prepared from HEK293 cells transfected with lacZ-overexpressing plasmid at 15 µg
Secondary
HRP-conjugated Goat anti-rabbit at 1/5000 dilution
developed using the ECL technique
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
Additional bands at : 90-100 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 2 seconds
Image courtesy of an anonymous Abreview.
References for Anti-beta Galactosidase antibody (ab616)
This product has been referenced in:
- Watkins Met al. Osteoblast connexin43 modulates skeletal architecture by regulating both arms of bone remodeling. Mol Biol Cell 22:1240-51 (2011).Read more (PubMed: 21346198) »
- Nazarenko Iet al. Brain abnormalities and glioma-like lesions in mice overexpressing the long isoform of PDGF-A in astrocytic cells. PLoS One 6:e18303 (2011).Read more (PubMed: 21490965) »
See all 53 publications for this product
Publishing research using ab616? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"