- PA5-19489 - Invitrogen Antibodies TUBA1C antibody

Submitted references Fourier Ring Correlation and Anisotropic Kernel Density Estimation Improve Deep Learning Based SMLM Reconstruction of Microtubules.

Berberich A, Kurz A, Reinhard S, Paul TJ, Burd PR, Sauer M, Kollmannsberger P
Frontiers in bioinformatics 2021;1:752788

A cytoskeletal function for PBRM1 reading methylated microtubules.

Karki M, Jangid RK, Anish R, Seervai RNH, Bertocchio JP, Hotta T, Msaouel P, Jung SY, Grimm SL, Coarfa C, Weissman BE, Ohi R, Verhey KJ, Hodges HC, Burggren W, Dere R, Park IY, Prasad BVV, Rathmell WK, Walker CL, Tripathi DN
Science advances 2021 Apr;7(14)

The MiDAC histone deacetylase complex is essential for embryonic development and has a unique multivalent structure.

Turnbull RE, Fairall L, Saleh A, Kelsall E, Morris KL, Ragan TJ, Savva CG, Chandru A, Millard CJ, Makarova OV, Smith CJ, Roseman AM, Fry AM, Cowley SM, Schwabe JWR
Nature communications 2020 Jun 26;11(1):3252

Ensemble-Level Organization of Human Kinetochores and Evidence for Distinct Tension and Attachment Sensors.

Roscioli E, Germanova TE, Smith CA, Embacher PA, Erent M, Thompson AI, Burroughs NJ, McAinsh AD
Cell reports 2020 Apr 28;31(4):107535

Mitotic spindle association of TACC3 requires Aurora-A-dependent stabilization of a cryptic α-helix.

Burgess SG, Mukherjee M, Sabir S, Joseph N, Gutiérrez-Caballero C, Richards MW, Huguenin-Dezot N, Chin JW, Kennedy EJ, Pfuhl M, Royle SJ, Gergely F, Bayliss R
The EMBO journal 2018 Apr 13;37(8)

Senescence-associated ultrastructural features of long-term cultures of induced pluripotent stem cells (iPSCs).

Colasuonno F, Borghi R, Niceforo A, Muzzi M, Bertini E, Di Giulio A, Moreno S, Compagnucci C
Aging 2017 Oct 23;9(10):2209-2222

FGFR3-TACC3 cancer gene fusions cause mitotic defects by removal of endogenous TACC3 from the mitotic spindle.

Sarkar S, Ryan EL, Royle SJ
Open biology 2017 Aug;7(8)

The mesh is a network of microtubule connectors that stabilizes individual kinetochore fibers of the mitotic spindle.

Nixon FM, Gutiérrez-Caballero C, Hood FE, Booth DG, Prior IA, Royle SJ
eLife 2015 Jun 19;4

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of acetylated alpha-Tubulin was performed using whole cell lysates from cells left untreated (DMSO only) or cells treated with 0.3uM or 3uM Trichostatin A (TSA) for 16 hours. Antigen-antibody complexes were formed by incubating 500 µg of lysate with 3 µg of an Acetyl Lysine monoclonal antibody (Product # MA1-2021) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with an alpha-Tubulin polyclonal antibody (Product # PA5-19489) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with Clean-blot IP Detection Reagent (Product # 21230) at a dilution of 1:2000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34087).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of alpha Tubulin was performed using 70% confluent log phase LNCaP 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of HeLa cells using Product # PA5-19489, anti-Alpha Tubulin antibody. The cells were fixed with methanol (100%) for 5 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. Partial rescue of mitotic defects in RT112 cells upon mild overexpression of TACC3. ( a ) Western blot analysis of TACC3 expression level from different promoters (pCMV versus pTACC3 or pPGK promoter) using alpha-GFP or alpha-TACC3 antibodies. alpha-Tubulin was used as the loading control. pCMV, promoter of cytomegalovirus; pTACC3, promoter of TACC3; pPGK, promoter of phosphoglycerate kinase. ( b ) Representative micrographs of the amount of GFP-TACC3 overexpression under pCMV and pTACC3. Scale bar, 10 um. ( c , d ) RT112 cells co-expressing H2B-mCherry with GFP or GFP-TACC3 under pTACC3, synchronized by a double thymidine block. Movies were started 3 h post-release and imaged every 3 min for the next 12 h. Proportions of cells with various mitotic phenotypes are shown ( c ). Categories are as described for all other figures. Cumulative histograms of mitotic progression of all cells ( d ). Prometaphase-metaphase timing is shown for the conditions indicated. GFP: n = 3, 111 cells. GFP-TACC3: n = 3, 95 cells. The full mitotic progression dataset is shown in the electronic supplementary material, figure S1.

PA5-19489

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