Anti-LOXL2 antibody (ab96233)
Key features and details
- Rabbit polyclonal to LOXL2
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-LOXL2 antibody
See all LOXL2 primary antibodies -
Description
Rabbit polyclonal to LOXL2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P MouseHumanWB Human
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Immunogen
Recombinant fragment corresponding to Human LOXL2. Recombinant fragment, corresponding to a region within amino acids 454-712 of Human LOXL2.
Database link: Q9Y4K0 -
Positive control
- IHC-P: Mouse ovary tissue and human esophageal cancer tissue. WB: Human samples only: MDA-MB-231 cell line, 293T, A431 and H1299 whole cell lysates, HeLa cell lysate. ICC/IF: A431 cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 78% PBS, 1% BSA, 20% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOXL2 antibody (ab96233)
Immunohistochemical analysis of Paraffin-embedded human esophageal carcinoma tissue labeling LOXL2 with ab96233 at 1/3000 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H2O2, RT, 30min. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Primary antibody incubation: 4°C overnight. Secondary antibody incubation: ABC HRP Kit (Rabbit IgG), 1:200, RT, 30min. Washing condition: PBS, 2 x 5 mins. Chromogen system: DAB.
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Western blot - Anti-LOXL2 antibody (ab96233)All lanes : Anti-LOXL2 antibody (ab96233) at 1/500 dilution (4? overnight)
Lane 1 : WT HeLa cell lysate at 30 µg
Lane 2 : LOXL2 knockout (KO) HeLa cell lysate
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 87 kDa7.5% SDS-PAGE.
alpha Tubulin was used as a loading control and showed equal loading.
Running: 80V, 15min; 140V, 40 min. Transfer: Semi-dry, 18 V, 60 min (NC embrane). Blocking: 5% non-fat milk in TBST, RT, 60 minutes. Washing: 5 ml TBST, 4 x 5 min. ECL exposure.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOXL2 antibody (ab96233)Immunohistochemical analysis of paraffin-embedded mouse ovary tissue labeling LOXL2 with ab96233 at 1/500 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H2O2, RT, 30min. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Primary antibody incubation: 4°C overnight. Secondary antibody incubation: ABC HRP Kit (Rabbit IgG), 1:200, RT, 30min. Washing condition: PBS, 2 x 5 mins. Chromogen system: DAB.
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Western blot - Anti-LOXL2 antibody (ab96233)All lanes : Anti-LOXL2 antibody (ab96233) at 1/1000 dilution
Lane 1 : MCF7 cell line
Lane 2 : MDA-MB-231 cell line
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 87 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?MCF-7 lack LOXL2 expression (PMID: 19330836, PMID:12154058 and PMID: 27655685).
7.5% gel. Running: 80V, 15min; 140V, 40 min. Transfer: Semi-dry, 18 V, 60 min (Nitrocellulose membrane). Blocking: 5% non-fat milk in TBST, RT, 60 min. Primary antibody incubation: 4°C overnight. Washing: 5 ml TBST, 4 x 5 min. Exposure: Western blot HRP substrate.
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Immunocytochemistry/ Immunofluorescence - Anti-LOXL2 antibody (ab96233)Immunocytochemistry/ Immunofluorescence analysis of A431 cells labeling LOXL2 with ab96233 at 1:500 (green). Cells were fixed in ice cold methanol for 5 minutes. Cells were co-stained with Hoechst 33342 (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LOXL2 antibody (ab96233)IHC image of ab96233 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96233, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Western blot - Anti-LOXL2 antibody (ab96233)All lanes : Anti-LOXL2 antibody (ab96233) at 1/1000 dilution
Lane 1 : 293T cell lysate
Lane 2 : A431 cell lysate
Lane 3 : H1299 cell lysate
Lysates/proteins at 30 µg per lane.
Predicted band size: 87 kDa
7.5% SDS-PAGE
