Anti-alpha Tubulin antibody - Loading Control (ab4074)
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- Specific References (158)
- Protocols
Overview
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Product nameAnti-alpha Tubulin antibody - Loading Control
See all alpha Tubulin primary antibodies -
DescriptionRabbit polyclonal to alpha Tubulin - Loading Control
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Host speciesRabbit
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SpecificityThis antibody detects a single band in cell lines, corresponding to alpha tubulin.
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Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
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Species reactivityReacts with: Mouse, Rat, Chicken, Cow, Human
Predicted to work with: Pig, Xenopus laevis, Chimpanzee, Monkey, Zebrafish, Chinese hamster
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human alpha Tubulin.
(Peptide available as ab23537.)
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Positive control
- This antibody gave a positive signal in HeLa cells in WB and IF.
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General notes
This antibody makes an excellent loading control.
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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Primary antibody notesThis antibody makes an excellent loading control.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunohistochemistry kits
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Related Products
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 | Abreviews | Notes |
|---|---|---|
| IHC-P | Use at an assay dependent concentration. | |
| WB | Use 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). | |
| ICC/IF | Use a concentration of 1 µg/ml. |
Target
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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%").
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Cellular localizationMajor constituent of Microtubules
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Database links
- Entrez Gene: 7277 Human
- Entrez Gene: 7278 Human
- Entrez Gene: 7846 Human
- Entrez Gene: 22142 Mouse
- Entrez Gene: 64158 Rat
- Entrez Gene: 399313 Xenopus laevis
- Entrez Gene: 373080 Zebrafish
- Omim: 191110 Human
see all -
Alternative names
- TUBA1 antibody
- TUBA1A antibody
- TUBA2 antibody
see all
Images
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Western blot - Anti-alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
Lane 2 : NIH 3T3 (Mouse) Whole Cell Lysate (ab52956)
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab4074)Image from PLoS One. 2014; 9(10): e109590. Fig 5o,doi: 10.1371/journal.pone.0109590Immunocytochemistry of SisNPFR-C6E8 cell line. Cells were prefixed in freshly prepared 4% paraformaldehyde (PFA) in F12 K culture medium with 10% fetal bovine serum and Geneticin. Fixed cells were permeabilized in PBST (0.25% Triton X-100 in PBS) for 5 min and blocked at 4°C overnight in 5% normal goat serum. The cells were then incubated at 4°C overnight with ab4074 (1:50.) Cover slips were placed onto slides and mounted with VectaShield containing DAPI.
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Western blot - Anti-alpha Tubulin antibody - Loading Control (ab4074)All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
Lane 2 : HEK293 (Human) Whole Cell Lysate (ab52256)
Lane 3 :NIH 3T3 whole cell lysate (ab7179)
Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lanes 6-7 :NIH 3T3 whole cell lysate (ab7179) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957) with Human alpha Tubulin peptide (ab23537) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-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.
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Western blot - alpha Tubulin antibody - Loading Control (ab4074) -
Immunocytochemistry/ Immunofluorescence - Anti-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 - Anti-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
All lanes : Goat Anti-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 secondsab4074 cross-reacts strongly with Mouse 3T3 lysate and shows weak cross-reactivity with Rat liver, Cow, Chicken and CHO lysates.
Protocols
References
This product has been referenced in:
- Deng WT et al. Human L- and M-opsins restore M-cone function in a mouse model for human blue cone monochromacy. Mol Vis 24:17-28 (2018). Read more (PubMed: 29386880) »
- Moore KS et al. Csde1 binds transcripts involved in protein homeostasis and controls their expression in an erythroid cell line. Sci Rep 8:2628 (2018). WB . Read more (PubMed: 29422612) »
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