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Purified Mouse Anti-Adaptin α
Purified Mouse Anti-Adaptin α

Western blot analysis of Adaptin α on a rat cerebrum lysate (left). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the anti-Adaptin α antibody.

Immunofluorescent staining of SH-SY5Y cells (right).  Cells were seeded in a 384 well collagen coated imaging plate (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-Adaptin-α antibody.  The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™  855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software  This antibody also stained SK-N-SH, C6, U87 and U373 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).

Western blot analysis of Adaptin α on a rat cerebrum lysate (left). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the anti-Adaptin α antibody.

Immunofluorescent staining of SH-SY5Y cells (right).  Cells were seeded in a 384 well collagen coated imaging plate (Material # 353962) at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure; Bioimaging protocol link) and the anti-Adaptin-α antibody.  The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen)(pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). The image was taken on a BD Pathway™  855 or 435 Bioimager System using a 20x objective and merged using the BD AttoVison ™ software  This antibody also stained SK-N-SH, C6, U87 and U373 cells using both the Triton X100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).

Product Details
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BD Transduction Laboratories™
Rat (QC Testing), Human, Mouse, Dog (Tested in Development)
Mouse IgG1
Mouse Adaptin α [A] aa. 38-215
Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required), Immunoprecipitation (Tested During Development)
112 kDa
250 µg/ml
AB_397867
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
  5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Triton is a trademark of the Dow Chemical Company.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
610501 Rev. 2
Antibody Details
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8/Adaptin α

Sorting of integral membrane proteins at various stages of the endocytic and secretory pathways is mediated by vesicular trafficking between a variety of organelles. Two sorting signals are tyrosine-based and dileucine-based signals that interact with heterotetrameric adaptor protein complexes (AP-1, AP-2, AP-3, and AP-4), which are associated with the vesicle coats. These coatomers contain two large Adaptin proteins (α, γ, δ, or ε and β1, β2, β3, or β4, respectively) that are noncovalently linked to one medium chain (µ1, µ2, µ3, or µ4) and one small chain (σ1, σ2, σ3, or σ4). The AP-1 and AP-3 complexes are involved in protein sorting from the TGN and endosomes, while AP-2 adaptor complexes are involved in clathrin-mediated endocytosis. In the AP-2 complex, Adaptin α is expressed in two very similar isoforms from two different genes. Adaptin α [A] (112kDa) is expressed primarily in brain, while Adaptin α [C] (105kDa) is expressed in brain, liver and other tissues.

610501 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
610501 Rev.2
Citations & References
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Development References (5)

  1. Jarousse N, Kelly RB. The AP2 binding site of synaptotagmin 1 is not an internalization signal but a regulator of endocytosis. J Cell Biol. 2001; 154(4):857-866. (Biology: Western blot). View Reference
  2. Robinson MS. Cloning of cDNAs encoding two related 100-kD coated vesicle proteins (alpha-adaptins). J Cell Biol. 1989; 108(3):833-842. (Biology). View Reference
  3. Santolini E, Puri C, Salcini AE, et al. Numb is an endocytic protein. J Cell Biol. 2000; 151(6):1345-1351. (Biology: Immunoprecipitation, Western blot). View Reference
  4. Teo M, Tan L, Lim L, Manser E. The tyrosine kinase ACK1 associates with clathrin-coated vesicles through a binding motif shared by arrestin and other adaptors. J Biol Chem. 2001; 276(21):18392-18398. (Biology: Immunofluorescence, Western blot). View Reference
  5. Wasiak S, Legendre-Guillemin V, Puertollano R, et al. Enthoprotin: a novel clathrin-associated protein identified through subcellular proteomics. J Cell Biol. 2002; 158(5):855-862. (Biology: Immunofluorescence, Western blot). View Reference
View All (5) View Less
610501 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.