BD, 610499, BD Transduction Laboratories™ Purified Mouse Anti-Clathrin Heavy Chain

Reactivity: Human (QC Testing), Mouse, Rat, Dog, Chicken (Tested in Development)
Isotype: Mouse IgG1
Immunogen: Rat Clathrin Heavy Chain aa. 4-171
Application: Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry, Immunoprecipitation (Tested During Development)
Target Molecular Weight: 180 kDa
Concentration: 250 µg/ml
RRID: AB_397866
Storage Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent staining and bioimaging Methanol Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample. Triton-X 100 Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States. 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. Triton is a trademark of the Dow Chemical Company. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to http://regdocs.bd.com to access safety data sheets (SDS). Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.