beta Actin Mouse Monoclonal Antibody [A2-F6]
cat.: EM21002
| Product Type: | Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, 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: | 2ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 42 kDa |
| Isotype: | IgG1 |
| Immunogen: | Synthetic peptide (KLH-coupled) within human Beta-actin aa 1-50. |
| Positive control: | Hela cell lysate, THP-1 cell lysate, HepG2 cell lysate, MCF-7 cell lysate, PC-12 cell lysate, PMVEC cell lysate, RH-35 cell lysate, L6 cell lysate, NIH/3T3 cell lysate, L929 cell lysate, RAW264.7 cell lysate, B16F1 cell lysate, MEF cell lysate, Hela, A549, NIH/3T3, human colon carcinoma tissue, mouse prostate tissue, mouse kidney tissue. |
| Subcellular location: | Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:10,000-160,000 1:100-1:200 1:1,000 1:1,000 |
| 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 β-actin β actin |
Images
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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. |
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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. |
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Fig3:
Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (EM21002) at 1/20,000 dilution. Lane 1: THP-1 cell lysate Lane 2: HepG2 cell lysate Lane 3: PMVEC cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: L929 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 42 kDa Observed band size: 42 kDa Exposure time: 2 minutes; 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 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Immunocytochemistry analysis of HeLa cells labeling beta Actin with Mouse anti-beta Actin antibody (EM21002) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-beta Actin antibody (EM21002) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Mouse anti-beta Actin antibody (EM21002) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002) at 1/1,000 dilution for 1 hour 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Mouse anti-beta Actin antibody (EM21002) at 1/1,000 dilution. 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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002) at 1/1,000 dilution for 1 hour 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. |
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