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UBC12 Recombinant Mouse mAb

Cell line: Ramos Fixative: 4% Paraformaldehyde Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:200 Primary incubation condition: 4°C overnight Secondary Ab: Goat Anti-Mouse IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for AP94431
Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:1000 Primary Ab incubation condition: 4°C overnight Secondary Ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: 1: HeLa, 2: HEK-293, 3: 4T1, 4: Raw264.7 Protein loading quantity: 20 µg Exposure time: 30 s Predicted MW: 21 kDa Observed MW: 21 kDa
Tissue: Human liver cancer Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:100 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: SP Kit(Mouse)(sp-0024) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for AP94431
Tissue: Rat colon Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:100 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: SP Kit(Mouse)(sp-0024) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for AP94431
Tissue: Mouse kidney Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:100 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: SP Kit(Mouse)(sp-0024) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for AP94431

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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-P, IHC-F, IF, ICC
Host	Rabbit
Clonality	Recombinant
Physical State	Liquid
Isotype	IgG1, Kappa
Purity	affinity purified by Protein G
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SIMILARITY	Belongs to the ubiquitin-conjugating enzyme family. UBC12 subfamily.
SUBUNIT	Interacts with UBA3, DCUN1D1 and RBX1.
Post-translational modifications	The acetylation of Met-1 is cotranslational, and not regulatory. The N-acetylmethionine increases affinity for DCUN1D1 by about 2 orders of magnitude and is crucial for NEDD8 transfer to cullins.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	Accepts the ubiquitin-like protein NEDD8 from the UBA3-NAE1 E1 complex and catalyzes its covalent attachment to other proteins. The specific interaction with the E3 ubiquitin ligase RBX1, but not RBX2, suggests that the RBX1-UBE2M complex neddylates specific target proteins, such as CUL1, CUL2, CUL3 and CUL4. Involved in cell proliferation.

Additional Information
Dilution	WB=1:500-1:1000,IHC-P=1:100-500,IHC-F=1:100-500,ICC/IF=1:50-1:200,IF=0
Format	0.01M TBS(pH7.4) with 1% BSA, 0.09% (W/V) sodium azide and 50% Glyce
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol