Anti-alpha Tubulin antibody - Loading Control (ab89984)
Overview
- Product nameAnti-alpha Tubulin antibody - Loading ControlSee all alpha Tubulin primary antibodies ...
- DescriptionChicken polyclonal to alpha Tubulin - Loading Control
- Tested applicationsWB, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Monkey - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human alpha Tubulin.
(Peptide available as ab23537.)
- Positive controlThis antibody gave a positive signal in the following whole cell lysates: HeLa; NIH 3T3; PC12;
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab89984 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 50 kDa).Can be blocked with alpha Tubulin peptide (ab23537). |
| ICC/IF | ICC/IF: Use a concentration of 5 µg/ml. |
Target
- RelevanceMicrotubules are involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintenance of cell shape. Tubulin itself is a globular protein which consists of two polypeptides (alpha and beta tubulin). Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter. Tyrosine ligase adds a C-terminal tyrosin to monomeric alpha tubulin. Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated alpha tubulin is referred to as Glu-tubulin. Another post-translational modification of detyrosinated alpha tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle. Alpha tubulin is not suitable as a loading control in adipose tissue as expression of tubulin in adipose tissue is very low ( Spiegelman and Farmer, Cell, 1982, 29(1): 53-60, "in cells undergoing adipose differentiation actin synthesis decreases by 90%").
- Cellular localizationMajor constituent of Microtubules
-
Database links
- Entrez Gene: 429035 Chicken
- Entrez Gene: 504244 Cow
- Entrez Gene: 7277 Human
- Entrez Gene: 7846 Human
- Entrez Gene: 7278 Human
- Entrez Gene: 22142 Mouse
- Entrez Gene: 64158 Rat
- Omim: 191110 Human
- SwissProt: Q5E986 Cow
- SwissProt: P68366 Human
- SwissProt: Q71U36 Human
- SwissProt: Q13748 Human
- SwissProt: P68369 Mouse
- SwissProt: P68370 Rat
see all
-
Alternative names
- TUBA1 antibodyTUBA1A antibodyTUBA2 antibody
- TUBA3 antibodyTUBA3C antibodyTUBA4A antibodyTubulin, alpha 1a antibodyTubulin, alpha 3c antibodyTubulin, alpha 4a antibody
see all
Anti-alpha Tubulin antibody - Loading Control images
-
Western blot - alpha Tubulin antibody - Loading Control (ab89984)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab89984) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Chicken IgY - H&L (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 50 kDa
Observed band size : 53 kDa (why is the actual band size different from the predicted?)
Additional bands at : 125 kDa,37 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds -
Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody - Loading Control (ab89984)ICC/IF image of ab89984 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab89984 at 5ug overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti- chicken IgY (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in HepG2 PFA fixed cell types at 5ug/ml.
-
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab89984)
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 50 kDa
Exposure time : 30 seconds
References for Anti-alpha Tubulin antibody - Loading Control (ab89984)
This product has been referenced in:
- Bassi ZI et al. Sticky/Citron kinase maintains proper RhoA localization at the cleavage site during cytokinesis. J Cell Biol 195:595-603 (2011). Read more (PubMed: 22084308) »
