alpha Tubulin Antibody (PSH02-94)

Novus Biologicals | Catalog # NBP3-33139

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH02-94 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within Human alpha Tubulin aa 402-451 / 451. (Uniprot: P68363)

Localization

Cytoplasm, Cytoskeleton, Microtubule.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

55 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for alpha Tubulin Antibody (PSH02-94)

alpha Tubulin Antibody (PSH02-94)

Western Blot: alpha Tubulin Antibody (PSH02-94) [NBP3-33139]

Western Blot: alpha Tubulin Antibody (PSH02-94) [NBP3-33139] - Western blot analysis of alpha Tubulin on different lysates with Rabbit anti-alpha Tubulin antibody (NBP3-33139) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: PC-12 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: Daudi cell lysate
Lane 5: Jurkat cell lysate
Lane 6: A431 cell lysate
Lane 7: K-562 cell lysate

Lysates/proteins at 20 ug/Lane.

Predicted band size: 50 kDa
Observed band size: 50 kDa

Exposure time: 1 minutes 2 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-33139) at 1/5,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
alpha Tubulin Antibody (PSH02-94)

Immunohistochemistry: alpha Tubulin Antibody (PSH02-94) [NBP3-33139]

Immunohistochemistry: alpha Tubulin Antibody (PSH02-94) [NBP3-33139] - Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-alpha Tubulin antibody (NBP3-33139) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-33139) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
alpha Tubulin Antibody (PSH02-94)

Immunocytochemistry/ Immunofluorescence: alpha Tubulin Antibody (PSH02-94) [NBP3-33139]

Immunocytochemistry/ Immunofluorescence: alpha Tubulin Antibody (PSH02-94) [NBP3-33139] - Immunocytochemistry analysis of Daudi cells labeling alpha Tubulin with Rabbit anti-alpha Tubulin antibody (NBP3-33139) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-alpha Tubulin antibody (NBP3-33139) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
alpha Tubulin Antibody (PSH02-94)

Flow Cytometry: alpha Tubulin Antibody (PSH02-94) [NBP3-33139]

Flow Cytometry: alpha Tubulin Antibody (PSH02-94) [NBP3-33139] - Flow cytometric analysis of HeLa cells labeling alpha Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-33139, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Applications for alpha Tubulin Antibody (PSH02-94)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:250-1:500

Immunohistochemistry-Paraffin

1:1000

Western Blot

1:1000-1:5000

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Advanced Features

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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: alpha Tubulin

Tubulin is the major building block of microtubules. Microtubules function as structural and mobile elements in mitosis, intracellular transport, flagellar movement and in the cytoskeleton. The tubulin superfamily is composed of six distinct families. Genes from the alpha, beta and gamma tubulin families are found in all eukaryotes. Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulins. Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter with a theoretical molecular weight of 50 kDa.

Tyrosine ligase adds a C-terminal tyrosine 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.

Like GAPDH and beta-actin, alpha/beta tubulin is often used as a loading control in immunoblot applications (1). Alpha/beta tubulin is also good for counterstaining microtubules in immunofluorescence (2).

References

1. Hannen, R., Selmansberger, M., Hauswald, M., Pagenstecher, A., Nist, A., Stiewe, T.,... Bartsch, J. W. (2019). Comparative Transcriptomic Analysis of Temozolomide Resistant Primary GBM Stem-Like Cells and Recurrent GBM Identifies Up-Regulation of the Carbonic Anhydrase CA2 Gene as Resistance Factor. Cancers (Basel), 11(7). doi:10.3390/cancers11070921

2. Nel, M., Joubert, A. M., Dohle, W., Potter, B. V., & Theron, A. E. (2018). Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines. Drug Des Devel Ther, 12, 1881-1904. doi:10.2147/dddt.S152718

Long Name

Tubulin Alpha 1a

Alternate Names

Alpha-Tubulin 3, B-ALPHA-1, LIS3, TUBA1A, TUBA3, Tubulin B-Alpha-1

Gene Symbol

TUBA1A

Additional alpha Tubulin Products

Product Documents for alpha Tubulin Antibody (PSH02-94)

Certificate of Analysis

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Product Specific Notices for alpha Tubulin Antibody (PSH02-94)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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