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Pan Actin Recombinant Rabbit mAb

Cell line: Jurkat Fixative: 4% Paraformaldehyde Permeabilization: 90% Methanol Primary ab dilution: 1:100 Secondary ab: Goat anti Rabbit IgG Unlabelled control: The cell without incubation with primary antibody and secondary antibody (Black line). Isotype control: Rabbit monoclonal IgG (Blue line). Comment: Line red is the positive signal for AP94657
Cell line: NIH/3T3 Fixative: 4% Paraformaldehyde Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for AP94657
Cell line: HeLa Fixative: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for AP94657
Cell line: A431 Fixative: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for AP94657
Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:1000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: HeLa, Jurkat, HEK293, NIH/3T3, C2C12, Mouse brain, Rat brain Protein loading quantity: 20 µg Exposure time: 60 s Predicted MW: 42 kDa Observed MW: 42 kDa
Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Pan Actin) Monoclonal Antibody, Unconjugated (AP94657) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Pan Actin) Monoclonal Antibody, Unconjugated (AP94657) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

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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
Calculated MW	42 KDa
Physical State	Liquid
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Cytoskeleton.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.In vertebrates 3 main groups of actin isoforms, alpha, beta and gamma have been identified. The alpha actins are found in muscle tissues and are a major constituent of the contractile apparatus. The beta and gamma actins coexist in most cell types as components of the cytoskeleton and as mediators of internal cell motility.Actin proteins have very high homology. The immunogen for this product is based on the ACTC1 protein but this antibody will also recognize: ACTB, ACTA1, ACTG1, ACTA2, ACTG2.

Additional Information
Dilution	WB=1:500-1:1000,IHC-P=1:100-500,IHC-F=,ICC/IF=1:50,IF=0,Flow-Cyt=1:50-1:100
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