Product datasheet
Anti-PLEASE DO NOT USE THIS TARGET PAGE antibody Loading Control ab75186
2 Abreviews 7 References 4 Images
Overview
Product name
Anti-PLEASE DO NOT USE THIS TARGET PAGE antibody - Loading Control
Description
Rabbit polyclonal to PLEASE DO NOT USE THIS TARGET PAGE - Loading Control
Tested applications
WB, ICC/IF, IHC-P
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Synthetic peptide derived from within residues 1 - 100 of Human beta Actin.Read Abcam's
proprietary immunogen policy(Peptide available as ab13772.)
Positive control
This antibody gave a positive signal in the following whole cell lysates: Jurkat, A431, HEK293,
HepG2, MCF-7, NIH3T3 and PC12. ICC/IF: NIH3T3 cells IHC: Rat small intestine (FFPE)
General notes
ab75186 is raised using the same immunogen as ab8227. Batches sold under ab75186 have
passed QC testing in Western blot only. In our hands, this product did not work in ICC/IF. For
further information please contact our Customer Services team.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab75186 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
1
Application
Abreviews
WB
Notes
Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa
(predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin
peptide (ab13772).
ICC/IF
Use a concentration of 1 µg/ml.
IHC-P
Use a concentration of 0.5 µg/ml.
Target
Function
Actins are highly conserved proteins that are involved in various types of cell motility and are
ubiquitously expressed in all eukaryotic cells.
Involvement in disease
Defects in ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of
dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle
contraction, often leading to abnormal postures. DYTJ patients manifest progressive,
generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing
loss.
Sequence similarities
Belongs to the actin family.
Post-translational
modifications
ISGylated.
Cellular localization
Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated
mRNAs.
Anti-PLEASE DO NOT USE THIS TARGET PAGE antibody - Loading Control images
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IHC image of ab75186 staining beta Actin in
rat small intestine formalin fixed paraffin
embedded tissue sections, performed on a
Leica Bond. The section was pre-treated
using heat mediated antigen retrieval with
sodium citrate buffer (pH6, epitope retrieval
solution 1) for 20 mins. The section was then
incubated with ab75186, 0.5μg/ml dilution, for
15 mins at room temperature and detected
using an HRP conjugated compact polymer
system. DAB was used as the chromogen.
Immunohistochemistry (Paraffin-embedded
The section was then counterstained with
sections) - Anti-beta Actin antibody - Loading
haematoxylin and mounted with DPX. No
Control (ab75186)
primary antibody was used in the secondary
only control (shown on the inset).
For other IHC staining systems (automated
and non-automated) customers should
optimize variable parameters such as antigen
retrieval conditions, primary antibody
concentration and antibody incubation times.
ab75186 staining beta Actin in NIH3T3 cells.
The cells were fixed with 100% methanol
(5min), permeabilized with 0.1% Triton X-100
for 5 minutes and then blocked with 1%
BSA/10% normal goat serum/0.3M glycine in
0.1%PBS-Tween for 1h. The cells were then
incubated overnight at +4°C with ab75186 at
1μg/ml (shown in green) and ab195889,
Mouse monoclonal [DM1A] to alpha Tubulin Microtubule Marker (Alexa Fluor® 594), at
1/250 dilution (shown in red). Nuclear DNA
Immunocytochemistry/ Immunofluorescence -
was labelled with DAPI (shown in blue).
Anti-beta Actin antibody - Loading Control
Image was taken with a confocal microscope
(ab75186)
(Leica-Microsystems, TCS SP8).
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All lanes : Anti-PLEASE DO NOT USE THIS
TARGET PAGE antibody - Loading Control
(ab75186) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblastlike cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma
cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney
cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver
Western blot - beta Actin antibody - Loading
carcinoma cell line) Whole Cell Lysate
Control (ab75186)
Lane 5 : MCF7 (Human breast
adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : NIH 3T3 (Mouse) Whole Cell Lysate
(ab7179)
Lane 7 : PC12 (Rat adrenal
pheochromocytoma cell line) Whole Cell
Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 42 kDa
Observed band size : 42 kDa
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developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 42 kDa
Exposure time : 10 seconds
Western blot image using the Optiblot
Reducing Electrophoresis Kit - 10 x 10 cm (4Western blot - Anti-beta Actin antibody - Loading
20%) (ab119220) with the
Control (ab75186)
Prism Ultra Protein Ladder (ab116028) 5µl
used. We recommend using our ECL
substrate kit (ab65623) .
20ug of Lysate per lane and detection using
ab75186 diluted 1/250.
Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK293 cell lysate
Lane 5: HepG2 cell lysate
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Replacement or refund for products not performing as stated on the datasheet
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Terms and conditions
Guarantee only valid for products bought direct from Abcam or one of our authorized distributors
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