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Anti-Clathrin heavy chain Rabbit Monoclonal Antibody

Immunofluorescent analysis using the Antibody at 1:50 dilution.
Immunofluorescent analysis using the Antibody at 1:150 dilution.
Figure 1. Western blot analysis of CLTC using anti-CLTC antibody (M03134).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human PC-3 whole cell lysates.
Lane 5: rat brain tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CLTC antigen affinity purified monoclonal antibody (Catalog # M03134) at 1:1000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CLTC at approximately 192 kDa. The expected band size for CLTC is at 192 kDa.
Figure 2. IHC analysis of CLTC using anti-CLTC antibody (M03134).
CLTC was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CLTC Antibody (M03134) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 3. IHC analysis of CLTC using anti-CLTC antibody (M03134).
CLTC was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CLTC Antibody (M03134) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 4. IHC analysis of CLTC using anti-CLTC antibody (M03134).
CLTC was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CLTC Antibody (M03134) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 5. IHC analysis of CLTC using anti-CLTC antibody (M03134).
CLTC was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CLTC Antibody (M03134) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC
Primary Accession	Q00610/P53675
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Clathrin heavy chain Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Calculated MW	191615 Da
Application Details	WB 1:1000-1:5000 IHC 1:50-1:200 ICC/IF 1:50-1:200
Subcellular Localization	Cytoplasmic vesicle membrane ; Peripheral membrane protein ; Cytoplasmic side. Membrane, coated pit ; Peripheral membrane protein ; Cytoplasmic side. Melanosome. Cytoplasmic face of coated pits and vesicles. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: ABDH-3
Immunogen	A synthesized peptide derived from human Clathrin heavy chain
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.