rabbit polyclonal anti integrin β4  (Cell Signaling Technology Inc)

Journal: Fukushima Journal of Medical Science

Article Title: Quantitative analysis of β1,6GlcNAc-branched N -glycans on β4 integrin in cutaneous squamous cell carcinoma

doi: 10.5387/fms.2020-12

Figure Lengend Snippet: Increased β1,6GlcNAc-branched N -glycans on β4 integrin in cutaneous squamous cell carcinoma tissues. Relative amounts of β4 integrin and β1,6 GlcNAc-branched N -glycans on β4 integrin in cell lysates from human normal skin ( n = 1) and cutaneous squamous cell carcinoma tissues ( n = 5) were determined by ELISA as shown in Figure 1. The graph indicates the ratio of β1,6 GlcNAc-branched N -glycans to a β4 integrin in each sample. One-way ANOVA and Bonferroni post-test, mean ± SEM of three independent assays conducted in triplicate. *** P < 0.001 vs. normal skin.

Article Snippet: The wells were blocked with 200 μL of 1% BSA in PBS for 1 h at 37°C, followed by incubation with 100 μL of anti-rabbit polyclonal Ab against β4 integrin (H-101, Santa Cruz Biotechnology, #sc-9090, 1 : 500) in TBS-T for 1 h at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Mutual localization of the α6 and β4 integrin subunit and a presence of α6β4 heterodimer revealed by SIM and PLA. ( a ) α6 (red) and β4 (green) are localized in the plasma membrane overlying the acrosomal cap (for details see panel c , white arrows), apical hook and equatorial segment in an intact sperm head. ( b ) PLA confirmed the presence of the α6β4 heterodimer in depicted locations ( a ). ( c ) SIM dual-colour imaging showing merged image of α6 (red) and β4 (green) localized in the plasma membrane (white arrows). ( d ) Huygens software was used for a better visualization of the colocalization area (yellow). Colocalization maps are based on Pearson’s correlation coefficient. Nucleus is visualized with Dapi (blue). Scale bar represents 1 µm ( a – c ) and 2 µm ( d ).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Imaging, Software

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Gene expression of β4 integrin in the cell-type fractions. Cq value of the gene is normalized by reference gene Rps2 . Numbers > 1 are considered as strongly expressed in the individual cell-types.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: ( a ) Primers designing and ( b ) agarose gel electrophoresis of PCR products involving in the cytoplasmic domain of β4 integrin. 1–5 PCR products amplified by primer pairs (pp) in mRNA sperm samples after elutriation.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Agarose Gel Electrophoresis, Amplification

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Western blot immunodetection of the β4 integrin in protein extract from mouse epididymal sperm with mouse monoclonal anti-β4 integrin (sc-13543) antibody; (1) antibody reaction in protein extract from mouse sperm, (2) negative control with mouse IgG; detection of 200 kDa protein band corresponds to β4 integrin (black arrow), possible high molecular weight isoforms (white arrows), non-specific reaction (grey arrows).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Western Blot, Immunodetection, Negative Control, Molecular Weight

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Mutual localization of α3 and β1 integrin subunit and a presence of α3β1 heterodimer revealed by SIM and PLA. ( a ) α3 (green) and β1 (red) are localized in the acrosomal cap area of intact sperm head, ( b ) PLA confirmed a presence of the α3β1 heterodimer. ( c ) SIM depicted their mutual localization in same structures. ( d ) Huygens software was used for better visualization of colocalization area (yellow) of α3 and β1. Colocalization maps are based on Pearson’s correlation coefficient. Nucleus is visualized with Dapi (blue). Scale bar represents 1 μm ( a – c ) and 2 μm ( d ).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Software

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Docking of α/β integrin N-terminal domains. ( a , b ) An example of resting state structure of extracellular domains of the α3β1 integrin as obtained from I-TASSER homology modelling. As expected from the crystal structures available for threading by I-TASSER the overall architecture of the resulting model is similar to αXβ2 (PDB ID 4neh; ) or αIIbβ3 (PDB ID 3fcs; ) heterodimer structures. The interacting region of N-terminal domains is highlighted by the black rectangle. Panel ( a ) shows the α3 subunit on top of the β1 domains, while panel ( b ) represents the same structure rotated by 90 degrees orienting both the N-terminal domains to the front of the image. ( c – f ) The predicted arrangement of heterodimers formed by N-terminal domains of integrins α3β1 ( c ), α6β1 ( d ), α3β4 ( e ), and α6β4 ( f ) from the flexible side chain docking of the domains by ClusPro. In the case of α3β4 ( f ) two binding modes are shown. The predicted most stable dimer with the β4 domain (orange) interacting through the NV residues homologous with the typical Arg/Lys finger of β domains is compared to an alternative, less stable binding mode, employing the RPEK sequence (grey). Both binding modes are similar and suggest that the α3β4 complex adopts different domain orientation compared to the remaining α3β1, α6β1, and α6β4 cases. The α3 domains are shown in green, β1 in blue, α6 in red, and β4 in orange.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Binding Assay, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Graphical summary of integrin heterodimers and their localization in intact sperm head. ( a ) Localization of integrin heterodimers as follows: α3β1, α6β1, α6β4–plasma membrane (PM) overlaying the apical acrosome (AA), α6β1, α6β4–sperm hook (H); α3β–outer acrosomal membrane of apical acrosome (AA); α6β4–inner acrosomal membrane and equatorial segment (ES) up to posterior post-acrosomal region (PAR). ( b ) Schematic depiction of integrin subunits anchored in the membrane, dynamics between α3 and α6 with β1 and β subunits interaction with cytoskeleton.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Model of activated states of α/β integrin heterodimers. A probable arrangement of integrin heterodimers in the open state was derived from the N-terminal domain complexes predicted by docking (see ) by a structural superposition of the existing crystal structure of αIIBβ3 in the open state (PDB ID 3fcu; ). The modelling suggests that while for the α3β1, α6β1 ( a ), and α6β4 ( b ) the integrins are involved in a cis interaction adopting the expected conformation with the membrane proximal domains separated [ , , , ], the orientation of the α3β4 N-terminal domains would lead to a complex with α and β subunits pointing in nearly opposite directions ( b ) supporting the possible trans interaction (see for more details).

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Addressing the Compartmentalization of Specific Integrin Heterodimers in Mouse Sperm

doi: 10.3390/ijms20051004

Figure Lengend Snippet: Diagram of sperm specific integrin subunits and heterodimers. The solid lines show proven integrin heterodimers α3β1, α6β1 and α6β4; the arrows show predicted α3 and α6 dynamics with β1; α3 and β4 cis interaction are energetically unstable in contract to trans interaction. Heterodimers between αV and β1/β3 are not known.

Article Snippet: Sperm were blocked with 10% BSA in PBS for 1 h and incubated with primary rabbit polyclonal antibody anti-β1 integrin (sc-8978, Santa Cruz Biotechnology, Inc.) diluted 1:10 in PBS and/or primary goat polyclonal antibody anti-α3 integrin (N19) (sc-6588, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) 1:10, mouse monoclonal antibody anti α6 integrin (F6) (sc-374057, Santa Cruz Biotechnology, Inc.) 1:10, rabbit polyclonal antibody anti-β4 integrin (bs-4115R, Bioss antibodies, Woburn, MA, USA) 1:10 in PBS over night at 4 °C, followed by Alexa Fluor 488 goat anti-rabbit IgG and/or Alexa Fluor 568 donkey anti-goat IgG (Molecular Probes, Eugene, OR, USA), Alexa Fluor 568 donkey anti-mouse IgG (Molecular Probes) secondary antibodies 1:300 in PBS for 1 h at room temperature.

Techniques:

Journal: PLoS ONE

Article Title: Regional Regulation of Purkinje Cell Dendritic Spines by Integrins and Eph/Ephrins

doi: 10.1371/journal.pone.0158558

Figure Lengend Snippet: Effect of integrin and Eph activation on the number and the intensity of pFAK- positive spines in Purkinje cells.

Article Snippet: Antibodies used in Purkinje cell experiment are: mouse monoclonal anti Calbindin (Swant 1/2000), guinea pig polyclonal anti calbindin (Synaptic System), chicken polyclonal anti calbindin (Swant 1/2000), rabbit polyclonal anti integrin β1 (Chemicon AB1952 1/500), rabbit polyclonal anti integrin β3 (Chemicon 1/500), rabbit polyclonal anti integrin β4 (Chemicon 1/500), rabbit polyclonal anti integrin α3 extracellular domain Ralph3.2 (Santa Cruz), rabbit polyclonal anti integrin α3 cytoplasmic domain (Chemicon AB1920 1/500), rabbit polyclonal anti integrin α5 extracellular domain (Chemicon AB1921 1/500), rabbit polyclonal anti integrin α5 cytoplasmic domain (Chemicon AB1928 1/500), rabbit polyclonal anti integrin αV (Chemicon), mouse monoclonal anti PSD95 (Affinity Bioreagents MA1-046, 1/500), rabbit polyclonal anti VGluT1 (Synaptic System), guinea pig polyclonal anti VGluT2 (Synaptic System), rabbit polyclonal anti pFAK (Santa Sruz), MAP2, Alexa fluor-conjugated secondary antibodies series were all purchased from Invitrogen and used at a dilution of 1/1000.

Techniques: Activation Assay

Journal: PLoS ONE

Article Title: Regional Regulation of Purkinje Cell Dendritic Spines by Integrins and Eph/Ephrins

doi: 10.1371/journal.pone.0158558

Figure Lengend Snippet: Effect of integrin activation on length and density of spines in Purkinje cells.

Article Snippet: Antibodies used in Purkinje cell experiment are: mouse monoclonal anti Calbindin (Swant 1/2000), guinea pig polyclonal anti calbindin (Synaptic System), chicken polyclonal anti calbindin (Swant 1/2000), rabbit polyclonal anti integrin β1 (Chemicon AB1952 1/500), rabbit polyclonal anti integrin β3 (Chemicon 1/500), rabbit polyclonal anti integrin β4 (Chemicon 1/500), rabbit polyclonal anti integrin α3 extracellular domain Ralph3.2 (Santa Cruz), rabbit polyclonal anti integrin α3 cytoplasmic domain (Chemicon AB1920 1/500), rabbit polyclonal anti integrin α5 extracellular domain (Chemicon AB1921 1/500), rabbit polyclonal anti integrin α5 cytoplasmic domain (Chemicon AB1928 1/500), rabbit polyclonal anti integrin αV (Chemicon), mouse monoclonal anti PSD95 (Affinity Bioreagents MA1-046, 1/500), rabbit polyclonal anti VGluT1 (Synaptic System), guinea pig polyclonal anti VGluT2 (Synaptic System), rabbit polyclonal anti pFAK (Santa Sruz), MAP2, Alexa fluor-conjugated secondary antibodies series were all purchased from Invitrogen and used at a dilution of 1/1000.

Techniques: Activation Assay

Journal: PLoS ONE

Article Title: Regional Regulation of Purkinje Cell Dendritic Spines by Integrins and Eph/Ephrins

doi: 10.1371/journal.pone.0158558

Figure Lengend Snippet: Immunostaining against integrin α3 and α5 in Purkinje cells in culture (A-C) showing that integrins are expressed on the dendritic tree and in synapses where they are contiguous with the presynaptic marker bassoon (A, arrowhead). The presence of integrins in dendritic spines was further demonstrated by subcellular fractionation (E). IF = initial homogenate, P = spun down pellet, IP4 = synaptoneurosome fraction. β1 integrin was concentrated in the 1P4 fraction, which was also enriched with PSD-95 (D-E).

Article Snippet: Antibodies used in Purkinje cell experiment are: mouse monoclonal anti Calbindin (Swant 1/2000), guinea pig polyclonal anti calbindin (Synaptic System), chicken polyclonal anti calbindin (Swant 1/2000), rabbit polyclonal anti integrin β1 (Chemicon AB1952 1/500), rabbit polyclonal anti integrin β3 (Chemicon 1/500), rabbit polyclonal anti integrin β4 (Chemicon 1/500), rabbit polyclonal anti integrin α3 extracellular domain Ralph3.2 (Santa Cruz), rabbit polyclonal anti integrin α3 cytoplasmic domain (Chemicon AB1920 1/500), rabbit polyclonal anti integrin α5 extracellular domain (Chemicon AB1921 1/500), rabbit polyclonal anti integrin α5 cytoplasmic domain (Chemicon AB1928 1/500), rabbit polyclonal anti integrin αV (Chemicon), mouse monoclonal anti PSD95 (Affinity Bioreagents MA1-046, 1/500), rabbit polyclonal anti VGluT1 (Synaptic System), guinea pig polyclonal anti VGluT2 (Synaptic System), rabbit polyclonal anti pFAK (Santa Sruz), MAP2, Alexa fluor-conjugated secondary antibodies series were all purchased from Invitrogen and used at a dilution of 1/1000.

Techniques: Immunostaining, Marker, Fractionation

Journal: PLoS ONE

Article Title: Regional Regulation of Purkinje Cell Dendritic Spines by Integrins and Eph/Ephrins

doi: 10.1371/journal.pone.0158558

Figure Lengend Snippet: A. Quantitation of the effects of the various interactions on the percentage of spines positive for pFAK. B quantitation of effects on pFAK intensity. C quantitation of spine density. D quantitation of spine length. Treatment with 500μM of Mn2+ for 1 hour (B1-6) increased pFAK but had no effect on spine density. Treatment with EphrinA3 (C1-6) decreased the density of proximal spines and of integrin signaling, whereas combined treatment of Mn2+ and EphrinA3 (D1-6) restored both spine density and pFAK intensity to values closed to control. Finally, treatment with RGD peptide (E1-6) decreased pFAK intensity and spine length without affecting spine density.

Article Snippet: Antibodies used in Purkinje cell experiment are: mouse monoclonal anti Calbindin (Swant 1/2000), guinea pig polyclonal anti calbindin (Synaptic System), chicken polyclonal anti calbindin (Swant 1/2000), rabbit polyclonal anti integrin β1 (Chemicon AB1952 1/500), rabbit polyclonal anti integrin β3 (Chemicon 1/500), rabbit polyclonal anti integrin β4 (Chemicon 1/500), rabbit polyclonal anti integrin α3 extracellular domain Ralph3.2 (Santa Cruz), rabbit polyclonal anti integrin α3 cytoplasmic domain (Chemicon AB1920 1/500), rabbit polyclonal anti integrin α5 extracellular domain (Chemicon AB1921 1/500), rabbit polyclonal anti integrin α5 cytoplasmic domain (Chemicon AB1928 1/500), rabbit polyclonal anti integrin αV (Chemicon), mouse monoclonal anti PSD95 (Affinity Bioreagents MA1-046, 1/500), rabbit polyclonal anti VGluT1 (Synaptic System), guinea pig polyclonal anti VGluT2 (Synaptic System), rabbit polyclonal anti pFAK (Santa Sruz), MAP2, Alexa fluor-conjugated secondary antibodies series were all purchased from Invitrogen and used at a dilution of 1/1000.

Techniques: Quantitation Assay