HA600032

Pan-Actin Mouse Monoclonal Antibody [A2E1]

cat.: HA600032

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	A2E1

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.2% BSA, 50% 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 corresponding to N terminal of Human Alpha actin (smooth muscle, skeletal muscle, cardiac muscle).
Positive control:	A431 cell lysate, MCF7 cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse smooth muscle tissue lysate, rat heart tissue lysate, HeLa, human heart tissue, human skeletal muscle tissue, mouse heart tissue, mouse skeletal muscle tissue, rat heart tissue, rat skeletal muscle tissue.
Subcellular location:	Cytoplasm, Cytoskeleton
Recommended Dilutions: WB IHC-P IF-Cell	1:1,000 1:10,000 1:100
Uniprot #:	SwissProt: P68133 Human \| P68032 Human \| P62736 Human \| P68134 Mouse \| P68033 Mouse \| P62737 Mouse \| P68136 Rat \| P68035 Rat \| P62738 Rat
Alternative names:	a actin ACTA ACTA1 Actin, alpha skeletal muscle ACTS_HUMAN Alpha actin 1 Alpha-actin-1 ASMA MPFD AAT6 ACTA_HUMAN ACTA2 Actin alpha 2 smooth muscle aorta Actin aortic smooth muscle Actin, aortic smooth muscle ACTSA ACTVS Alpha 2 actin Alpha actin 2 Alpha cardiac actin alpha sma Alpha-actin-2 Cell growth inhibiting gene 46 protein Cell growth-inhibiting gene 46 protein GIG46 Growth inhibiting gene 46 MYMY5 ACTC ACTC_HUMAN ACTC1 Actin alpha cardiac muscle 1 Actin alpha cardiac muscle 1 Alpha-cardiac actin ASD5 CMD1R CMH11 LVNC4

Images

Fig1: Western blot analysis of Pan-Actin on different lysates with Mouse anti-Pan-Actin antibody (HA600032) at 1/1,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: HEK-293 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (20 µg/Lane)
Lane 6: Mouse smooth muscle tissue lysate (40 µg/Lane)
Lane 7: Rat heart tissue lysate (40 µg/Lane)

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

Exposure time: 9 seconds;

4-20% 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 (HA600032) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HeLa cells labeling Pan-Actin with Mouse anti-Pan-Actin antibody (HA600032) at 1/100 dilution.

Cells were fixed in 100% precooled methanol 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-Pan-Actin antibody (HA600032) 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) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human heart tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Mouse anti-Pan-Actin antibody (HA600032) at 1/10,000 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 (HA600032) at 1/10,000 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.