Product nameAnti-Drebrin antibody
See all Drebrin primary antibodies
DescriptionRabbit polyclonal to Drebrin
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, IPmore details
Species reactivityReacts with: Mouse, Rat, Chicken, Human, Quail
Predicted to work with: Cow
Synthetic peptide corresponding to Human Drebrin aa 600 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in the following lysates: SHSY-5Y; SK N SH; SK N BE; Human Cortex Neuronal cells; Rat Cortex Neuronal cells; Mouse Cortex Neuronal cells; Rat Hippocampus Tissue; Mouse Hippocampus Tissue; Malme 3M. ICC/IF: PC12 cells.
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab60933 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 71 kDa).|
|IHC-P||1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration.|
FunctionDrebrins might play some role in cell migration, extension of neuronal processes and plasticity of dendrites, respectively.
Tissue specificityBrain neurons. Also found in the heart, placenta, skeletal muscle, kidney and pancreas.
Sequence similaritiesContains 1 ADF-H domain.
- Information by UniProt
- D0S117E antibody
- DBN 1 antibody
- DBN1 antibody
ab60933 staining Drebrin in mouse brain tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using Citrate buffer pH6. Samples were then blocked with 1% BSA for 10 minutes at 21°C followed by incubation with the primary antibody at a 1/500 dilution, for 2 hours at 21°C. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as secondary antibody at a 1/200 dilution.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Drebrin knockout HAP1 whole cell lysate (20 µg)
Lane 3: SH-SY5Y whole cell lysate (20 µg)
Lane 4: Hu cerebellum whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab60933 observed at 100120 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab60933 was shown to specifically react with Drebrin in wild-type HAP1 cells as signal was lost in Drebrin knockout cells. Wild-type and Drebrin knockout samples were subjected to SDS-PAGE. ab60933 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-Drebrin antibody (ab60933) at 1 µg/ml
Lane 1 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 2 : SK N SH (Human neuroblastoma) Whole Cell Lysate
Lane 3 : SK N BE (Human neuroblastoma) Whole Cell Lysate
Lane 4 : Human Cortex Neuronal cell lysate
Lane 5 : Rat Cortex Neuronal cell lysate
Lane 6 : Mouse Cortex Neuronal cell lysate
Lane 7 : Rat Hippocampus Tissue Lysate
Lane 8 : Mouse Hippocampus Tissue Lysate
Lane 9 : Malme 3M (Human melanoma cells) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : 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: 71 kDa
Observed band size: 100,120 kDa why is the actual band size different from the predicted?
Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
The banding pattern observed is consistent with what has been described in the literature (PMID:10234022, 8929425). Drebrin has two isoforms produced by alternative splicing.
Drebrin was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to Drebrin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, SHSY5Y whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab60933.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 100kDa: Drebrin.
ICC/IF image of ab60933 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab60933, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) PC12 cells at 5µg/ml.
This product has been referenced in:
- Kommaddi RP et al. Aß mediates F-actin disassembly in dendritic spines leading to cognitive deficits in Alzheimer's disease. J Neurosci 38:1085-1099 (2018). Read more (PubMed: 29246925) »
- Wahl D et al. Comparing the Effects of Low-Protein and High-Carbohydrate Diets and Caloric Restriction on Brain Aging in Mice. Cell Rep 25:2234-2243.e6 (2018). Read more (PubMed: 30463018) »