Anti-alpha Tubulin antibody - Loading Control (ab4074)
- Datasheet
- References (207)
- 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
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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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Isotype control
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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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FunctionTubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
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Sequence similaritiesBelongs to the tubulin family.
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Post-translational
modificationsSome glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling. -
Cellular localizationCytoplasm > cytoskeleton.
- Information by UniProt
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Database links
- Entrez Gene: 7277 Human
- Entrez Gene: 22145 Mouse
- Entrez Gene: 316531 Rat
- Entrez Gene: 573122 Zebrafish
- Entrez Gene: 573216 Zebrafish
- Omim: 191110 Human
- SwissProt: P68366 Human
- SwissProt: P68368 Mouse
see all -
Alternative names
- Alpha-tubulin 1 antibody
- ALS22 antibody
- B ALPHA 1 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:
- Li W et al. Berberine suppresses IL-33-induced inflammatory responses in mast cells by inactivating NF-?B and p38 signaling. Int Immunopharmacol 66:82-90 (2019). Read more (PubMed: 30445310) »
- Yousefzadeh MJ et al. Mouse Models of Accelerated Cellular Senescence. Methods Mol Biol 1896:203-230 (2019). Read more (PubMed: 30474850) »
