PA5-30380

antibody from Invitrogen Antibodies

Targeting: TUBB4B Beta2, TUBB2C

Antibody data

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Validation data

Product number: PA5-30380 - Provider product page
Provider: Invitrogen Antibodies
Product name: Tubulin beta-2C Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: 293T, A431, Jurkat, Raji, NIH-3T3. Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (99%), Xenopus laevis (99%), Chicken (100%), Rhesus Monkey (100%), Chimpanzee (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1.66 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

TUBB4B protein structure - PA5-30380 shown in red.

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), PC-12 (Lane 6) and Neuro-2a (Lane 7). The blot was probed with Anti-Tubulin beta-2C Rabbit Polyclonal Antibody (Product # PA5-30380, 1:4000 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 Tubulin beta-2C was observed across the cell lines 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: Western Blot analysis of Tubulin-beta-2C was performed by separating 30 µg of A431 lysates by 10% SDS PAGE. Proteins were transferred to a membrane and probed with a Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380) at a dilution of 1:10000. 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 of Tubulin-beta-2C was performed by separating 30 µg of NIH-3T3 lysates by 10% SDS PAGE. Proteins were transferred to a membrane and probed with a Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380) at a dilution of 1:10000. 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 using Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380) diluted at 1:10,000. 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: Immunofluorescent analysis of beta Tululin 2C in methanol-fixed A431 cells using a beta Tululin 2C polyclonal antibody (Product # PA5-30380) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Tubulin beta-2C was performed using 70% confluent log phase SH-SY5Y 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 Tubulin beta-2C Rabbit Polyclonal Antibody (Product # PA5-30380) 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: Immunocytochemistry-Immunofluorescence analysis of Tubulin-beta-2C was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Tubulin beta-2C Polyclonal Antibody detects beta Tubulin 2C protein at cytoskeleton by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: beta Tubulin 2C stained by Tubulin beta-2C Polyclonal Antibody (Product # PA5-30380) diluted at 1:500. Blue: Fluoroshield with DAPI . Scale bar= 10 µm.