Overview

  • Product name
    Anti-Drebrin antibody [M2F6]
    See all Drebrin primary antibodies
  • Description
    Mouse monoclonal [M2F6] to Drebrin
  • Host species
    Mouse
  • Specificity
    This antibody detects Drebrin A and E in samples from human, mouse, rat, guinea pig, and cat origins and Drebrin E only in chicken samples.
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Fr, ICC, ICC/IF, IHC-FoFr, IHC-FrFl, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Guinea pig, Cat, Monkey, Quail
  • Immunogen

    Chicken Drebrin E protein.

  • Positive control
    • Mouse Brain Tissue Extract (ab7934).
  • General notes
    For maximum product recovery, after thawing, centrifuge the product vial before removing cap.

Properties

Applications

Our Abpromise guarantee covers the use of ab12350 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
WB Use a concentration of 10 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 65 kDa). The difference in the size of the detected band (~110 kDa) and in the predicted (~65 kDa) is due to the large number of negatively charged residues in this protein. Block membrane with 5-10% milk for 1 hour.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration. Fix cells with 4% PFA for 15 minutes. Permeabilise cells for 15 minutes with 0.1% triton X100 in PBS (see Ikeda K et al).
ICC/IF Use at an assay dependent concentration. See Toda et al.
IHC-FoFr 1/1000. See Keon et al.
IHC-FrFl 1/1000.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

Images

  • All lanes : Anti-Drebrin antibody [M2F6] (ab12350) at 1/5000 dilution

    Lanes 1-4 : Trangenic (APP/PS1) male mouse cortical lysate
    Lanes 5-8 : Wild-type male mouse cortical lysate
    Lane 9 : Rat cortical lysate

    Secondary
    All lanes : Alexa-Fluor 680 conjugated goat anti-mouse IgG

    Predicted band size: 65 kDa
    Observed band size: 120,42,95 kDa
    why is the actual band size different from the predicted?



    Western blot showing detection of drebrin in mouse and rat cortical lysates by ab12350.

    See Abreview

  • IHC-P image of Drebrin staining on Human ovarian carcinoma sections using ab12350 (1:100). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab12350 was diluted 1:100 in TBS buffer (containing BSA and Azide) and sections were then incubated with ab12350 for 16 hours at 21°C. The secondary antibody used was Biotin conjugated Goat polyclonal to Rabbit IgG (1:250).

    See Abreview

  • ab12350 at 1/50 staining mouse brain tissue sections by IHC-P. The tissue fixed in 4% Formalin, diluted in Dulbecco A PBS with standard pwax processing. Citric acid 0.01M pH6 was used as antigen retrieval solution. The tissue sections were incubated with the antibody for 2 hours. A biotinylated goat polyclonal antibody was used as the secondary. The image shows the lateral ventricle showing high cytoplasmic positivity in migrating progenitor neurones in the subventricular space.There is a low level of positivity in the dendritic field of the striatum; this appears to be specific. An identical pattern of staining is seen in same area of rat brain. All non-migratory cells in the same field are negative. The Corpus Callosum is negative.

    See Abreview

  • Immunohistochemistical detection (on formaldehyde/PFA-fixed paraffin-embedded sections) of Drebrin antibody [M2F6] (ab12350) on Quail Tissue sections (embryo d5/6 brain). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody: ab12350 incubated at 1/200 for 2 hours at RT. Secondary Antibody: Biotin labelled goat anti mouse Igs (1/200).
  • Immunofluorescent staining in the rat brain cerebellum with ab12350 mouse monoclonal Drebrin antibody. Drebrin-positive neurons and fibers are observed. Picture taken with X20 objective. Protocol: IHC free-floating protocol using 4% PFA fixed brain tissue. Rats were intracardially perfused with 4% PFA. Tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. Primary antibody ab12350 was used at 1/1000 incubated overnight at room temperature. Secondary antibody was Alexa Fluor 488 used at 1/1000, 2h incubation at room temperature. Image recoloured in Adobe photoshop.
  • Overlay histogram showing SH-SY5Y cells stained with ab12350 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab12350, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Western blot analysis of Drebrin expression in mouse brain (1) NIH/3T3 (2), Jurkat (3), HeLa (4), and PC12 (5) using Drebrin mAb (ab12350).

References

This product has been referenced in:
See all 25 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Rat Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Ovarian Carcinoma)
Specification
Ovarian Carcinoma
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted May 31 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT-116)
Loading amount
30 µg
Specification
HCT-116
Gel Running Conditions
Non-reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Mr. Qifeng Lin

Verified customer

Submitted Jan 13 2011

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Neural stem cell line, Cor-1)
Specification
Neural stem cell line, Cor-1
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Trition
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 28 2009

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neural stem cell line, Cor-1)
Loading amount
100 µg
Specification
Neural stem cell line, Cor-1
Gel Running Conditions
Reduced Denaturing (7% polyacrylamide gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Ms. Sangeetha Gajendra

Verified customer

Submitted Jul 02 2009

Application
Western blot
Sample
Mouse Tissue lysate - whole (brain)
Loading amount
30 µg
Specification
brain
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Aug 22 2008

Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12)
Loading amount
30 µg
Specification
C2C12
Gel Running Conditions
Reduced Denaturing (gradiant 4-20%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 19 2008

Application
Western blot
Sample
Mouse Tissue lysate - whole (Cortical lysates)
Loading amount
10 µg
Specification
Cortical lysates
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Martin Broadstock

Verified customer

Submitted May 02 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Quail Tissue sections (Quail embryo d5/6 brain)
Specification
Quail embryo d5/6 brain
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C

Mr. Carl Hobbs

Verified customer

Submitted Dec 04 2007

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Chicken Tissue sections (Developing chick head, E6)
Specification
Developing chick head, E6
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C

Mr. Carl Hobbs

Verified customer

Submitted Nov 19 2007

1-10 of 11 Abreviews

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