Anti-alpha Tubulin antibody - Loading Control (ab4074)
Overview
- Product nameAnti-alpha Tubulin antibody - Loading ControlSee all alpha Tubulin primary antibodies ...
- DescriptionRabbit polyclonal to alpha Tubulin - Loading Control
- SpecificityThis antibody detects a single band in cell lines, corresponding to alpha tubulin.
- Tested applicationsIHC-P, WB, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Chicken, Cow, Human
Predicted to work with: Pig, Xenopus laevis, Chimpanzee, Monkey, Zebrafish, Chinese Hamster - 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 HeLa cells in WB and IF.
- General notes
This antibody makes an excellent loading control.
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
- Primary antibody notes This antibody makes an excellent loading control.
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab4074 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| IHC-P | |
| WB | |
| ICC/IF |
IHC-P: 1/25.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). Can be blocked with alpha Tubulin peptide (ab23537).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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
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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
- Entrez Gene: 399313 Xenopus laevis
- Entrez Gene: 373080 Zebrafish
- Omim: 191110 Human
- SwissProt: Q5E986 Cow
- SwissProt: P68366 Human
- SwissProt: Q71U36 Human
- SwissProt: Q13748 Human
- SwissProt: P68369 Mouse
- SwissProt: P68370 Rat
- SwissProt: Q4V850 Xenopus laevis
- SwissProt: O42271 Zebrafish
see all
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Alternative names
- Alpha-tubulin 1 antibodyFLJ30169 antibodyH2 ALPHA antibody
- Testis specific alpha tubulin antibodyTUBA1 antibodyTUBA1A antibodyTUBA2 antibodyTUBA3 antibodyTUBA3C antibodyTUBA4A antibodyTubulin alpha 1 chain antibodyTubulin alpha antibodyTubulin H2 alpha antibodyTubulin, alpha 1a antibodyTubulin, alpha 3c antibodyTubulin, alpha 4a antibody
see all
Anti-alpha Tubulin antibody - Loading Control images
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Western blot - alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : Mouse 3T3 lysate
Lane 2 : Rat liver lysate
Lane 3 : Cow cell lysate
Lane 4 : Chicken cell lysate
Lane 5 : CHO lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 50 kDa
Observed band size : 50 kDa
Exposure time : 30 seconds -
Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody - Loading Control (ab4074)ICC/IF image of ab4074 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4074, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red). -
Western blot - alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 : HeLa whole cell lysate withalpha Tubulin peptide (ab23537) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Predicted band size : 50 kDa
Observed band size : 50 kDa -
Immunohistochemistry (Paraffin-embedded sections) - alpha Tubulin antibody - Loading Control (ab4074)(Image courtesy of Human Protein Atlas).
ab4074 staining alpha tubulin in the glandular cells of the endometrium. Paraffin embedded endometrium tissue of a 33 year-old female was incubated with ab4074 (1/25 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.ab4074 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org. -
Western blot - alpha Tubulin antibody - Loading Control (ab4074)Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml + whole cell lysate prepared from MCF7 cells at 20 µg
Secondary
Goat anti-rabbit IgG-HRP at 1/1000 dilution
developed using the ECL technique
Predicted band size : 50 kDa
Observed band size : 50 kDa
Exposure time : 5 minutesImage courtesy of an anonymous Abreview.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab4074)Image courtesy of an anonymous Abreview.ab4074 staining alpha Tubulin in murine sperm by Immunocytochemistry/ Immunofluorescence.Cells were fixed in 2% paraformaldehyde, permeabilized in -20°C methanol for 10 minutes, blocked using 3% BSA for 1 hour at 25°C and then incubated with ab4074 at a 1/300 dilution for 2 hours at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit (H+L) polyclonal used at a1/400 dilution.
References for Anti-alpha Tubulin antibody - Loading Control (ab4074)
This product has been referenced in:
- Pfeiffer JR & Brooks SA Cullin 4B is recruited to tristetraprolin-containing messenger ribonucleoproteins and regulates TNF-a mRNA polysome loading. J Immunol 188:1828-39 (2012). Read more (PubMed: 22262661) »
- Raiz J et al. The non-autonomous retrotransposon SVA is trans-mobilized by the human LINE-1 protein machinery. Nucleic Acids Res : (2011). WB ; Human . Read more (PubMed: 22053090) »
