AP2B1 antibody (AA 75-245)

Catalog No. ABIN967931

Product Details anti-AP2B1 Antibody

Target: AP2B1 (Adaptor-Related Protein Complex 2, beta 1 Subunit (AP2B1))

Binding Specificity: AA 75-245

Reactivity: Human, Mouse, Rat, Dog, Chicken, Frog

Host: Mouse

Clonality: Monoclonal

Conjugate: This AP2B1 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), BioImaging (BI)

Cross-Reactivity: Mouse (Murine), Rat (Rattus), Dog (Canine), Chicken, Frog

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. 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.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. 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.
5. Please refer to us for technical protocols.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Adaptin beta aa. 75-245

Clone: 74-Adaptin beta

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968537, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-AP2B1 antibody (ABIN967931)

Naga Prasad, Laporte, Chamberlain, Caron, Barak, Rockman: "Phosphoinositide 3-kinase regulates beta2-adrenergic receptor endocytosis by AP-2 recruitment to the receptor/beta-arrestin complex." in: The Journal of cell biology, Vol. 158, Issue 3, pp. 563-75, (2002) (PubMed).

Ros-Baro, Lopez-Iglesias, Peiro, Bellido, Palacin, Zorzano, Camps: "Lipid rafts are required for GLUT4 internalization in adipose cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, Issue 21, pp. 12050-5, (2001) (PubMed).

Laporte, Oakley, Zhang, Holt, Ferguson, Caron, Barak: "The beta2-adrenergic receptor/betaarrestin complex recruits the clathrin adaptor AP-2 during endocytosis." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, Issue 7, pp. 3712-7, (1999) (PubMed).

Ponnambalam, Robinson, Jackson, Peiperl, Parham: "Conservation and diversity in families of coated vesicle adaptins." in: The Journal of biological chemistry, Vol. 265, Issue 9, pp. 4814-20, (1990) (PubMed).

Kirchhausen, Nathanson, Matsui, Vaisberg, Chow, Burne, Keen, Davis: "Structural and functional division into two domains of the large (100- to 115-kDa) chains of the clathrin-associated protein complex AP-2." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 86, Issue 8, pp. 2612-6, (1989) (PubMed).

Target Details for AP2B1

Target: AP2B1 (Adaptor-Related Protein Complex 2, beta 1 Subunit (AP2B1))

Alternative Name: Adaptin beta (AP2B1 Products)

Background: 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 (gamma, alpha, delta, or epsilon and beta1, beta2, beta3, or beta4, respectively) that are noncovalently linked to one medium chain (µ1, µ2, µ3, or µ4) and one small chain (sigma1, sigma2, sigma3, or sigma4). 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. beta Adaptin subunits (beta1, beta2, beta3, beta4) lack sequence homology to adaptins alpha, gamma, delta, and epsilon, but all of these subunits share a similar domain structure. Adaptin beta1 (also known as Adaptin beta') and beta2 (also known as Adaptin beta) have 83% amino acid identity and are found in the AP1 and AP2 complexes, respectively.

Molecular Weight: 106 kDa

Pathways: EGFR Signaling Pathway, Neurotrophin Signaling Pathway, EGFR Downregulation