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ACTA1 antibody - N-terminal region (ARP60969_P050)

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100 ul

Regular Price: $289.00

Special Price: $215.00

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Conjugation Options

ARP60969_P050-FITC Conjugated

ARP60969_P050-HRP Conjugated

ARP60969_P050-Biotin Conjugated

Gene Symbol:
ACTA1
NCBI Gene Id:
58
Official Gene Full Name:
Actin, alpha 1, skeletal muscle
Protein Name:
Actin, alpha skeletal muscle
Swissprot Id:
P68133
Protein Accession #:
NP_001091
Nucleotide Accession #:
NM_001100
Alias Symbols:
ACTA, ASMA, CFTD, CFTD1, CFTDM, MPFD, NEM1, NEM2, NEM3
Replacement Item:
This antibody may replace item sc-10731 from Santa Cruz Biotechnology.
Description of Target:
The product encoded by this gene belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actins being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. This actin is an alpha actin that is found in skeletal muscle. Mutations in this gene cause nemaline myopathy type 3, congenital myopathy with excess of thin myofilaments, congenital myopathy with cores, and congenital myopathy with fiber-type disproportion, diseases that lead to muscle fiber defects.
Protein Size (# AA):
377
Molecular Weight:
42kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express ACTA1.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express ACTA1.
Species Reactivity:
Cow, Guinea Pig, Horse, Human, Rabbit, Rat, Mouse
Predicted Homology Based on Immunogen Sequence:
Cow: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Rabbit: 100%; Rat: 100%
Complete computational species homology data:
Anti-ACTA1 (ARP60969_P050)
Peptide Sequence:
Synthetic peptide located within the following region: GQKDSYVGDEAQSKRGILTLKYPIEHGIITNWDDMEKIWHHTFYNELRVA
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
ABL1; TRIM63; TRIM55; CEP57; AMOT; UBC; SUMO3; CDKN2A; ACTG1; BRK1; ACTBL2; NCKAP1; LRCH3; MAPK14; DSTN; NMI; UBL4A; ABRA; ABLIM2; POLR1B; RRN3; HTRA2; PTPN22; ABLIM3; BTK; CCT2; TRAF3IP1; TCP1; SNCA; PARK2; LGALS3; SHC1; GSN; CDK2; CUL1; ISG15; YWHAZ; RE
Blocking Peptide:
For anti-ACTA1 (ARP60969_P050) antibody is Catalog # AAP60969 (Previous Catalog # AAPP47112)
Datasheets/Manuals:
Printable datasheet for anti-ACTA1 (ARP60969_P050) antibody
 
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2 Item(s)

02/01/2017 16:24
  • Overall Experience:
  • Quality:
Product Review: ACTA1 antibody - N-terminal region (ARP60969_P050) in Mouse GL261 cell and astrocyte cell using Western blot
Product page for ACTA1 antibody - N-terminal region (ARP60969_P050)

Researcher: Anonymous
Application: Western blotting
Species + Tissue/Cell type: Lane 1: 20ug mouse GL261 cell lysate Lane 2: 20ug mouse astrocyte cell lysate
Primary antibody dilution: 1:500
Secondary antibody: Anti-rabbit-HRP
Secondary antibody dilution: 1:500


Questionnaire:
How do Aviva's reagents play a role in your experimental goals? Use as a loading control for WB
How would you rate this antibody on a scale from 1-5 (5=best) and why? 4 - close to good
Would you use this antibody in future experiments? May be
Have you used another antibody which has worked in your application? Yes
Do you believe the information about the reagent on Aviva's website is correct? Yes (but I used lower dilution)
How did you store the antibody after re-suspension?  -20C
Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel): Human; Glial cells
How many different experimental trials were conducted using the antibody sample? 2
How was this sample prepared? Cell cultures in 60 mm dishes were scraped into lysis buffer (PBS with a cocktail of protease inhibitors (Sigma) plus 1% Triton X-100), incubated for 30 min. at 4C with constant rotation, and centrifuged at 10,000 rpm for 10 min.
Primary antibody dilution and incubation time:  1:500
Secondary antibody used and dilution and incubation time:  1:500
What controls were used in your experiment (positive/negative)? It was control
Please include your detailed WB Procedure/Protocol here: Western blotting.
1. Cell cultures in 60 mm dishes were scraped into lysis buffer (PBS with a cocktail of protease inhibitors (Sigma) plus 1% Triton X-100), incubated for 30 min. at 4C with constant rotation, and centrifuged at 10,000 rpm for 10 min. The protein concentration was measured with Bio-Rad Protein assay (#500-0006).
2. Forty micrograms of protein was separated by electrophoresis in 10% sodium dodecyl sulfate-polyacrylamide gel and then transferred to polyvinylidene difluoride membrane (Bio-Rad#162-0177) at 60V for 3h in cold room (4C).
3. The blots were blocked in 5% nonfat milk (Bio-Rad #170-6404) prepared in PBS + 0.1% Tween-20 for 1 h at room temperature (RT) with constant shaking.
4.  Then the membranes were incubated with Primary antibodies o/n (~18h) at 4C with constant rotation.
5. After washing (shaking 3 times for 5 min each in PBS+0.1% Tween-20) the Secondary antibodies (horseradish peroxidase-conjugated anti-rabbit IgG (Thermo Scientific/Pierce#1858415)) were applied at dilution 1:500 - 1:1,000 for 1 h at RT with constant rotation.
6. After washing (3 times for 5 min each) the signals were detected with SuperSignal West Pico (or Femto)  Chemiluminescent Substrate (Thermo Scientific/Pierce #34077 or #34095).
7. All blots were reprobed for b-actin to control the protein loading. At first, the membranes were stripped with Restore PLUS Western Blot Stripping Buffer (Thermo Scientific#46430), then blocked in 5% milk (see step 3), and then incubated with anti-Actin Ab (1:500) o/n at 4C, washed, and incubated with  horseradish peroxidase-conjugated anti-rabbit IgG (1:500) followed washing and ECL development.
We used CL-XPosure Film (Thermo Scientific#34090).
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02/01/2017 16:24
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Product Review: ACTA1 antibody - N-terminal region (ARP60969_P050) in Human PC3 prostate cancer cells using Western Blot
Product Page for ACTA1 antibody - N-terminal region (ARP60969_P050)

Researcher: Dr. Hafiz Ahmed, University of Maryland School of Medicine and Greenebaum Cancer Center
Application: Western blotting
Species + Tissue/Cell type: Human PC3 prostate cancer cells
How many ug's of tissue/cell lysate run on the gel:
1. 50 ug human PC3 cell lysate
2. 50 ug human PC3 cell lysate (Serum Starved)
Primary antibody dilution: 1:1000
Secondary antibody: Goat anti-Rabbit HRP
Secondary antibody dilution: 1:5000



Questionnaire:

How do Aviva's reagents play a role in your experimental goals?

Actin antibody was used as loading control.

How would you rate this antibody on a scale from 1-5 (5=best) and why?

4 (background is minimum).

Would you use this antibody in future experiments?

Yes.

Have you used another antibody which has worked in your application?

Not yet.

Do you believe the information about the reagent on Aviva's website is correct?

Yes.

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?

Yes, we will publish data.

How did you store the antibody after re-suspension?

Aliquoted to small fractions and store at -20 oC. Used the aliquot for each experiment.

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):

Human PC3 prostate cancer cells, 50 ug protein was loaded on each lane.

How many different experimental trials were conducted using the antibody sample?

About 10 times.

How was this sample prepared?

Used Cell lysis kit (Sigma).

Primary antibody dilution and incubation time:

1 ug/ml.

Secondary antibody used and dilution and incubation time:

Goat anti-rabbit IgG-HRP.

What controls were used in your experiment (positive/negative)?

Usually galectin-3 antibody as a positive control.

No primary antibody, but only secondary as a negative control.

Please include your detailed WB Procedure/Protocol here:

Cells were extracted in cell lysis buffer (Sigma) and after centrifugation protein was estimated. Loaded equal amount of protein in each lane and perfomed SDS-PAGE. The protein bands were transferred to nirocellulose membrane (Western blot), blocked with 1% BSA dissolved in PBS/Tween 20 (0.1%), and incubated over night with the primary antibody (in this case, the anti-actin antibody described above) at 4 oC.  The membrane was washed 3 times with PBS/Tween 20 (20 min each) and incubated with secondary antibody in PBS/Tween 20 containing 0.1% BSA for 1 h at room temperature. After washing the blot as above, chemiluminiscence substrate was added for 5 min and the image was captured in X-Ray film.
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