The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml.
Use a concentration of 3 µg/ml.
Use a concentration of 10 µg/ml.
Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
FunctionGTPase-activating protein for RhoA and Cdc42 small GTPases. Together with CDC42 seems to be involved in the pathway mediating the repulsive signaling of Robo and Slit proteins in neuronal migration. SLIT2, probably through interaction with ROBO1, increases the interaction of SRGAP1 with ROBO1 and inactivates CDC42.
Tissue specificityExpressed in brain, lung, kidney, and testis.
ICC/IF image of ab57504 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 ab57504 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 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.
Western blot - SRGAP1 antibody (ab57504)
SRGAP1 antibody (ab57504) at 1ug/lane + IMR-32 cell lysate at 25ug/lane.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SRGAP1 antibody (ab57504)This image is courtesy of an anonymous Abreview
ab57504 staining SRGAP1 in Human Fetal Brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized with 0.1% Triton-X-100 i PBS; antigen retrieval was by heat mediation with a citrate buffer (10mM). Samples were incubated with primary antibody (1/900 in 3% serum/0.1% Triton-X-100 in PBS) for 16 hours at 4°C. A Biotin-conjugated Horse anti-mouse polyclonal (1/200) was used as the secondary antibody.
Immunohistochemistry (Frozen sections) - SRGAP1 antibody (ab57504)Image courtesy of an anonymous Abreview.
ab57504 staining SRGAP1 in human fetal brainstem tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with paraformaldehyde and permeabilized using 0.1% Triton X-100/PBS. Samples were then incubated with 3% serum and the primary antibody at a 1/1000 dilution, for 16 hours at 4°C. A biotin-conjugated horse anti-mouse IgG polyclonal was used as the secondary antibody at a 1/200 dilution.
Overlay histogram showing SH-SY5Y cells stained with ab57504 (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 (ab57504, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/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 with 0.1% PBS-Tween for 20 min used under the same conditions.
References for Anti-SRGAP1 antibody (ab57504)
This product has been referenced in:
Chen ZB et al. Slit-Robo GTPase-Activating Proteins are Differentially Expressed in Murine Dorsal Root Ganglia: Modulation by Peripheral Nerve Injury. Anat Rec (Hoboken)295:652-60 (2012).
Read more (PubMed: 22271578) »
Ip BK et al. The corticofugal neuron-associated genes ROBO1, SRGAP1, and CTIP2 exhibit an anterior to posterior gradient of expression in early fetal human neocortex development. Cereb Cortex21:1395-407 (2011).
Read more (PubMed: 21060114) »