Immunofluorescent analysis of alpha Adaptin was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with alpha Adaptin (AC1-M11) Mouse Monoclonal Antibody (MA3-061) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Amphibian, Bovine, Chicken, Human, Mouse, Non-human primate, Pig, Rat|
|Published species reactivity||Rat, Non-human primate, Amphibian, Bovine, Mouse, Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified bovine brain adaptor complexes.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||2 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||2 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA3-061 detects alpha-adaptin from bovine, chicken, frog, human, monkey, mouse, pig and rat. This antibody detects the products of both alpha-adaptin genes, alphaA and alphaC, as well as an alternatively spliced isoform of alphaA found in neurons.
MA3-061 has been successfully used in Western blot, immunofluorescence, and immunoprecipitation procedures. By Western blot, this antibody recognizes an ~100 kDa doublet representing the two isoforms of alpha-adaptin from pig brain extract. Immunofluorescence staining of alpha-adaptin in transfected COS cells with MA3-061A yields distinct punctate dots on the plasma membrane.
The MA3-061 antigen is purified bovine brain adaptor complexes.
Clathrin-mediated endocytosis is the pathway by which many receptors for nutrients and hormones are internalized to be recycled or down-regulated. During formation of clathrin coated membranes, clathrin co-assembles with heterotetrameric molecules known as assembly polypeptides (APs) or adaptors which form a layer of protein between the clathrin lattice and the membrane.
There are two characterized adaptors AP1 and AP2. AP1 is associated with clathrin coated vesicles at the trans-Golgi network and AP2 is associated with the endocytic clathrin coated vesicles at the plasma membrane and has been shown to specifically interact with Src homologous and collagen-like (Shc) and epidermal growth factor (EGF) receptor. AP2 is composed of four subunits, two separate 100 kDa gene products with similar domain structures (alpha and beta adaptin) and a 50 and 17 kDa subunit. There are two alpha-adaptin genes, alphaA and alphaC which have a tissue specific pattern of expression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Clathrin and phosphatidylinositol-4,5-bisphosphate regulate autophagic lysosome reformation.
MA3-061 was used in western blot to study the regulation of autophagic lysosome reformation by clathrin and phosphatidylinositol-4,5-bisphosphate
|Rong Y,Liu M,Ma L,Du W,Zhang H,Tian Y,Cao Z,Li Y,Ren H,Zhang C,Li L,Chen S,Xi J,Yu L||Nature cell biology (14:924)||2012|
Recruitment of endophilin to clathrin-coated pit necks is required for efficient vesicle uncoating after fission.
MA3-061 was used in western blot to investigate the role of endophilin recruitment in vesicle uncoating during endocytosis
|Milosevic I,Giovedi S,Lou X,Raimondi A,Collesi C,Shen H,Paradise S,O'Toole E,Ferguson S,Cremona O,De Camilli P||Neuron (72:587)||2011|
Synaptic and vesicular coexistence of VGLUT and VGAT in selected excitatory and inhibitory synapses.
MA3-061 was used in western blot to investigate the colocalization of VGLUT and VGAT and its effect on neurotransmitter uptake
|Zander JF,Münster-Wandowski A,Brunk I,Pahner I,Gómez-Lira G,Heinemann U,Gutiérrez R,Laube G,Ahnert-Hilger G||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:7634)||2010|
Endocytosis in cultured neurons is altered by chronic alcohol exposure.
MA3-061 was used in western blot to investigate the influence of chronic alcohol consumption on neuronal endocytosis
|Marín MP,Esteban-Pretel G,Ponsoda X,Romero AM,Ballestín R,López C,Megías L,Timoneda J,Molowny A,Canales JJ,Renau-Piqueras J||Toxicological sciences : an official journal of the Society of Toxicology (115:202)||2010|
Silencing of fas-associated death domain protects mice from septic lung inflammation and apoptosis.
MA3-061 was used in western blot to investigate the effects of siRNA-mediated silencing of fas-associated death domain (FADD) on septic lung inflammation and apoptosis
|Matsuda N,Yamamoto S,Takano K,Kageyama S,Kurobe Y,Yoshihara Y,Takano Y,Hattori Y||American journal of respiratory and critical care medicine (179:806)||2009|
Xenopus autosomal recessive hypercholesterolemia protein couples lipoprotein receptors with the AP-2 complex in oocytes and embryos and is required for vitellogenesis.
MA3-061 was used in western blot to demonstrate the function of xARH in the interactions among LDL, vitellogenin receptors, and the AP-2 complex.
|Zhou Y,Zhang J,King ML||The Journal of biological chemistry (278:44584)||2003|
A beta-turn endocytic code is required for optimal internalization of the growth hormone receptor but not for alpha-adaptin association.
MA3-061 was used in western blot to study the mechanism for the internalization of the growth hormone receptor.
|Vleurick L,Pezet A,Kühn ER,Decuypere E,Edery M||Molecular endocrinology (Baltimore, Md.) (13:1823)||1999|
100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties.
MA3-061 was used in western blot to study coat protein membrane binding properties in the Golgi region
|Wong DH,Brodsky FM||The Journal of cell biology (117:1171)||1992|
ARAP1 regulates endocytosis of EGFR.
MA3-061 was used in immunocytochemistry to investigate the effect of ARAP1 on endocytosis of EGFR
|Yoon HY,Lee JS,Randazzo PA||Traffic (Copenhagen, Denmark) (9:2236)||2008|
A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis.
MA3-061 was used in immunocytochemistry to investigate the role of dynamin 1 in nervous system function
|Ferguson SM,Brasnjo G,Hayashi M,Wölfel M,Collesi C,Giovedi S,Raimondi A,Gong LW,Ariel P,Paradise S,O'toole E,Flavell R,Cremona O,Miesenböck G,Ryan TA,De Camilli P||Science (New York, N.Y.) (316:570)||2007|
Endocytosis of epithelial apical junctional proteins by a clathrin-mediated pathway into a unique storage compartment.
MA3-061 was used in immunocytochemistry to investigate the mechanism of endocytosis of adherens junction proteins and tight junction proteins in T84 epithelial cells
|Ivanov AI,Nusrat A,Parkos CA||Molecular biology of the cell (15:176)||2004|
Cytoplasmic transport of Stat3 by receptor-mediated endocytosis.
MA3-061 was used in immunocytochemistry to study the mechanism for the translocation of Stat3 through the cytoplasm to the nucleus
|Bild AH,Turkson J,Jove R||The EMBO journal (21:3255)||2002|
Interaction of SPIN90 with syndapin is implicated in clathrin-mediated endocytic pathway in fibroblasts.
MA3-061 was used in immunohistochemistry to investigate the interaction of SPIN90 and syndapin and its role in clathrin-mediated endocytic pathway
|Kim SH,Choi HJ,Lee KW,Hong NH,Sung BH,Choi KY,Kim SM,Chang S,Eom SH,Song WK||Genes to cells : devoted to molecular and cellular mechanisms (11:1197)||2006|