ET1701-80

Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]

cat.: ET1701-80

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IF-Cell, IF-Tissue, IHC-P
Clonality:	Monoclonal
Clone number:	JJ09-29

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 42 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Actin aa 45-80 / 377.
Positive control:	HeLa cell lysate, A431 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, mouse colon tissue lysate, rat brain tissue lysate, rat colon tissue lysate, human colon carcinoma tissue, hybrid fish (crucian-carp) brain tissue lysate, hybrid fish (crucian-carp) kidney tissue lysate, NIH/3T3, mouse cardiac muscle tissue, mouse smooth muscle tissue.
Subcellular location:	Cytoplasm, Cytoskeleton.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:5,000 1:100-1:500 1:100-1:500 1:50-1:200
Uniprot #:	SwissProt: P68133 Human \| P68134 Mouse \| P68136 Rat Entrez Gene: 407658 Zebrafish
Alternative names:	a actin ACTA ACTA1 Actin alpha skeletal muscle Actin actin, alpha 1, skeletal muscle 1 actin, alpha 1, skeletal muscle Actin, alpha skeletal muscle actina actine ACTS_HUMAN aktin Alpha Actin 1 Alpha skeletal muscle Actin alpha skeletal muscle alpha-actin Alpha-actin-1 ASMA CFTD CFTD1 CFTDM MPFD NEM1 NEM2 NEM3 nemaline myopathy type 3

Images

Fig1: Western blot analysis of Actin on different lysates with Rabbit anti-Actin antibody (ET1701-80) at 1/5,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: A431 cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: PC-12 cell lysate (15 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: Mouse colon tissue lysate (20 µg/Lane)
Lane 7: Rat brain tissue lysate (20 µg/Lane)
Lane 8: Rat colon tissue lysate (20 µg/Lane)

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

Exposure time: 24 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 (ET1701-80) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig2: Immunofluorescence analysis of paraffin-embedded human colon carcinoma tissue labeling Actin with Rabbit anti-Actin antibody (ET1701-80) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-80, green) at 1/100 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

	Fig3: Western blot analysis of 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 (ET1701-80, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hybrid fish (crucian-carp) brain tissue lysate Lane 2: Hybrid fish (crucian-carp) kidney tissue lysate
	Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling Actin with Rabbit anti-Actin antibody (ET1701-80) 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 Rabbit anti-Actin antibody (ET1701-80) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue using anti-Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-80, 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse smooth muscle tissue using anti-Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-80, 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”.