PA5-29546

antibody from Invitrogen Antibodies

Targeting: TUBA4A FLJ30169, H2-ALPHA, TUBA1

Antibody data

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Validation data

Product number: PA5-29546 - Provider product page
Provider: Invitrogen Antibodies
Product name: TUBA4A Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Application Note: For IHC, epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections.
Reactivity: Human, Mouse, Rat, Canine
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.44 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

TUBA4A protein structure - PA5-29546 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TUBA4A was performed by separating 30 µg of NIH-3T3 lysates by 10% SDS PAGE. Proteins were transferred to a membrane and probed with a TUBA4A Polyclonal Antibody (Product # PA5-29546) at a dilution of 1:3000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TUBA4A was performed by separating 30 µg of Hep G2 whole cell extracts by 10% SDS PAGE. Proteins were transferred to a membrane and probed with a TUBA4A Polyclonal Antibody (Product # PA5-29546) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using TUBA4A Polyclonal Antibody (Product # PA5-29546). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with TUBA4A Polyclonal Antibody (Product # PA5-29546) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), C2C12 (Lane 4), MDA-MB-231 (Lane 5), PC-12 (Lane 6), RSC96 (Lane 7) and tissue extracts of Mouse Lung (Lane 8). The blot was probed with Anti-TUBA4A Rabbit Polyclonal Antibody (Product # PA5-29546, 1:2000 dilution) 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: Immunofluorescent analysis of alpha Tubulin 4a in paraformaldehyde-fixed U2OS cells using an Alpha Tubulin 4a polyclonal antibody (Product # PA5-29546) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of TUBA4A in paraformaldehyde-fixed U2OS cells using TUBA4A Polyclonal Antibody (Product # PA5 29546) (Green) at a dilution of 1:500. Red: Alpha tubulin filaments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TUBA4A 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 TUBA4A Rabbit Polyclonal Antibody (Product # PA5-29546) 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.