Beta Actin Mouse Monoclonal Antibody [A2-F6]
cat.: EM21002
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, ICC, IHC-P, FC
Clonality: Monoclonal
Clone number: A2-F6
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2 mg/mL.
Purification: Protein A affinity purified.
Molecular weight: 42 kDa
Isotype: IgG1
Immunogen: Synthetic peptide (KLH-coupled) within human Beta-actin N terminal.
Positive control: NIH/3T3 cell lysate, PC-12 cell lysate, MCF-7 cell lysate, HepG2 cell lysate, Hela cell lysate, mouse lung tissue lysate, Hela, A549, NIH/3T3, human colon carcinoma tissue, mouse prostate tissue, mouse kidney tissue.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  ICC
  IHC-P
  FC

1:10,000-160,000
1:100-1:200
1:100-1:200
1:100-1:200
Uniprot #: SwissProt: P60709 Human | P60710 Mouse | P60711 Rat
Alternative names: A26C1A A26C1B ACTB ACTB_HUMAN Actin beta Actin cytoplasmic 1 Actin, cytoplasmic 1, N-terminally processed Actx b actin Beta cytoskeletal actin Beta-actin BRWS1 E430023M04Rik MGC128179 PS1TP5 binding protein 1 PS1TP5BP1
Images
EM21002_1.jpg Fig1: Western blot analysis of Beta Actin on Hela cell lysates with Mouse anti-Beta Actin antibody (EM21002).

Hela cell lysates at 10 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM21002) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
EM21002_2.jpg Fig2: Western blot analysis of Beta Actin on different lysates with Mouse anti-Beta Actin antibody (EM21002) at 1/40,000 dilution.

Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM21002) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
EM21002_3.jpg Fig3: Western blot analysis of Beta actin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM21002, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.

Positive control:
Lane 1: NIH/3T3 cell lysate, 10 µg/Lane
Lane 2: PC-12 cell lysate, 10 µg/Lane
Lane 3: MCF-7 cell lysate, 10 µg/Lane
Lane 4: HepG2 cell lysate, 10 µg/Lane
Lane 5: Hela cell lysate, 10 µg/Lane
Lane 6: Mouse lung tissue lysate, 20 µg/Lane
EM21002_4.jpg Fig4: ICC staining of Beta Actin in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
EM21002_5.jpg Fig5: ICC staining of Beta Actin in A549 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
EM21002_6.jpg Fig6: ICC staining of Beta Actin in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
EM21002_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Beta Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM21002_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse prostate tissue using anti-Beta Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM21002_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Beta Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.