Invitrogen

alpha Tubulin Polyclonal Antibody

Product Image Gallery

alpha Tubulin Antibody — Published figure using alpha Tubulin polyclonal antibody (Product # PA5-19489)

alpha Tubulin Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of alpha Tubulin was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Alpha Tubulin Rabbit Polyclonal Antibody (Product # PA5-19489) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

alpha Tubulin Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), NIH/3T3 (Lane 4), HT-29 (Lane 5), Neuro-2a (Lane 6) and tissue extract of Rat Brain (Lane 7). The blot was probed with Rabbit Anti-alpha Tubulin Polyclonal Antibody (Product # PA5-19489, 0.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 52 kDa band corresponding to alpha Tubulin was observed across the cell lines and tissue tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

0.01 µg/1x10^6 cells

Immunocytochemistry (ICC)

1-5 µg/mL

Immunofluorescence (IF)

1-5 µg/mL

Immunohistochemistry (Paraffin) (IHC (P))

1:2500

Western Blot (WB)

0.5 µg/mL

Published Applications

Immunofluorescence (IF)

See 1 publication below

Western Blot (WB)

See 2 publications below

Immunocytochemistry (ICC)

See 1 publication below

Product Specifications

Species Reactivity

Bovine, Dog, Chicken, Deer, Fruit fly, Hamster, Human, Mouse, Non-human primate, Rat

Published species

Human

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.

Conjugate

Unconjugated

Form

Liquid

Concentration

0.1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7.4, with 1% BSA

Contains

0.02% sodium azide

Storage conditions

-20°C or -80°C if preferred

RRID

AB_10984311

Product Specific Information

PA5-19489 targets alpha Tubulin in immunofluorescence, immunohistochemistry (paraffin), and Western blot applications and shows reactivity with chicken, Indian muntjac, Chinese hamster, human, fruit fly, rat, African Green Monkey, and mouse samples. Perform heat mediated antigen retrieval via the pressure cooker method before beginning immunohistochemistry staining protocol.

The PA5-19489 immunogen is synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.

PA5-19489 detects alpha Tubulin which has a predicted molecular weight of approximately 50 kDa.

Target Information

Alpha Tubulin is a part of a heterodimer consisting of alpha and beta tubulin subunits. The alpha and beta tubulins, which are each about 55 kDa MW, are homologous but not identical. Alpha-beta tubulin heterodimer is the basic building block of microtubules, and this intracellular cylindrical filamentous structure is present in almost eukaryotic cells. Microtubules serve as structural supports and lines of transport within the cell, as well as serving a key role in mitosis. Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulins. The alpha and beta tubulins represent the major components of microtubules, while gamma tubulin plays a critical role in the nucleation of microtubule assembly. There are multiple alpha and beta tubulin genes, which are highly conserved among species. The gene that encodes alpha tubulin is highly similar to the mouse and rat Tuba1 gene. Northern blotting studies have shown that the gene expression is predominantly found in morphologically differentiated neurologic cells.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: alpha tubulin; alpha tubulin 84B; alpha-Tubulin; Alpha-tubulin 1; Alpha-tubulin 4; alpha-Tubulin 84B; alpha-Tubulin at 84B; Alpha-tubulin isotype M-alpha-4; alpha-tubulin84B; alpha1 tubulin; alpha1-tubulin; alpha[[1]] tubulin; alpha[[1]]-tubulin; alphaT; alphaTub84B-PA; alphatubulin; alphaTubulin 84B; alphaTubulin84B; CG1913-PA; FLJ30169, H2-ALPHA, TUBA1, alpha-tubulin 1; Testis-specific alpha-tubulin; tubulin; tubulin 1 alpha-chain; tubulin 1alpha; Tubulin alpha-1 chain; Tubulin alpha-4 chain; Tubulin alpha-4A chain; tubulin alpha1; tubulin alpha1 promoter; Tubulin H2-alpha; tubulin(alpha1); tubulin, alpha 1; tubulin, alpha 1 (testis specific); tubulin, alpha 4; tubulin, alpha 4a; Tubulin-alpha1; tubulin1alpha; tubulin84B

Gene Aliases: 1t; a-tub84B; a1t; ALPHA 84C; alpha-Tub; alpha-Tub84B; alpha-tubulin; alpha-tubulin 84B; alpha1; alpha1t; alpha1tub; alpha1tub84B; alpha84B; alphaT; alphat-1; alphaTUB; alphaTub1; alphaTub84; alphaTUB84B; ALS22; anon-EST:Liang-1.59; anon-EST:Liang-2.30; aTub84B; BEST:LD32507; BOS_2117; CG1913; chr3R:2914276..2914416; clone 1.59; clone 2.30; DM1alpha; Dmel\CG1913; Dmel_CG1913; DTA1; H2-ALPHA; l(3)84Bd; l(3)g3; M[a]4; ms(3)nc33; nc33; T; Talpha1; TBA1_DROME; Tub; TUBA1; Tuba4; TUBA4A; Tuba84B; Tubalpha1; tubulin

UniProt ID: (Human) P68366, (Bovine) P81948, (Rat) Q5XIF6, (Mouse) P68368, (Fruit fly) P06603

Entrez Gene ID: (Dog) 478918, (Human) 7277, (Bovine) 777775, (Rat) 316531, (Chinese hamster) 100767686, (Mouse) 22145, (Fruit fly) 40848

Molecular Function: cytoskeletal protein microtubule family cytoskeletal protein tubulin