- PA5-16863 - Invitrogen Antibodies TUBB6 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot of Tubulin Beta using Tubulin Beta Polyclonal Antibody (Product # PA5-16863) on MCF-7 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30µg 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), tissue extracts of Mouse Lung (Lane 8), Rat Stomach (Lane 9) and Rat Brain (Lane 10). The blot was probed with Anti-beta Tubulin Rabbit Polyclonal Antibody (Product # PA5-16863, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). A 52 kDa band corresponding to beta Tubulin was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MCF-7 (Lane 2), A431 (Lane 3), A549 (Lane 4), HEK 293 (Lane 5) and HeLa (Lane6). The blots were probed with Anti-Tubulin Beta Rabbit Polyclonal Antibody (Product # PA5-16863, 1:1000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjµgate (Product # G-21234, 1:5000 dilution). A 50 kDa band corresponding to Tubulin was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX), 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 was performed on whole cell extracts (30µg 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), tissue extracts of Mouse Lung (Lane 8), Rat Stomach (Lane 9) and Rat Brain (Lane 10). The blot was probed with Anti-beta Tubulin Rabbit Polyclonal Antibody (Product # PA5-16863, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). A 52 kDa band corresponding to beta Tubulin was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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)
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Experimental details: Immunofluorescence analysis of Tubulin Beta 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 2% BSA for 1 hour at room temperature. The cells were labeled with Tubulin Beta Rabbit Polyclonal Antibody (Product # PA5-16863) 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) 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 Alexa Fluor® 555 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)
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Experimental details: Formalin-fixed, paraffin-embedded human lung stained with Tubulin beta antibody using peroxidase-conjugate and AEC. Note cytoplasmic staining of ciliated epithelium of bronchioles.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Tubulin Beta showing staining in the cytoplasm of paraffin-embedded human colon carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Tubulin Beta Rabbit Polyclonal Antibody (Product # PA5-16863) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Tubulin Beta showing staining in the cytoplasm of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Tubulin Beta Rabbit Polyclonal Antibody (Product # PA5-16863) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of Tubulin Beta was done on HeLa cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Tubulin Beta Rabbit Polyclonal Antibody (PA516863, red histogram) or with rabbit isotype control (yellow histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 6 Effect of inactivation of PMC1 on the level of Pmr1 in the ret1-27 mutant strain and in the strain bearing the RET1 wild-type allele. Proteins from cell lysates were resolved by SDS PAGE and transferred to nitrocellulose membrane, which was then divided in two parts at the level of the 80 kDa marker band. The upper part was stained with antiserum against H . polymorpha Pmr1, while the lower part was stained with antibody against tubulin used as a loading control. ret1-27 pmc1-Delta , the 64MA70QA-Deltapmc strain; pmc1-Delta , the 64MA70Q-RET-Deltapmc strain; ret1-27 , the 64MA70QAL strain; WT and WT 1/2, undiluted and two-fold diluted sample of the 64MA70QL-RET strain, respectively.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 SH3PXD2B silencing inhibits the invasion, but not proliferation of Hep3B and Huh7 cells. Hep3B and Huh7 cells were transduced with lentivirus for expression of SH3PXD2B-specific shRNA or control shRNA (Scr). The relative levels of SH3PXD2B to the control beta-tubulin protein expression in different groups of Hep3B ( a , b ) and Huh7 ( c , d ) cells were determined by Western blot. e The dynamic proliferation of Hep3B and Huh7 cells was determined at the indicated time points. f , g The invasion of different groups of Hep3B and Huh7 cells was examined by transwell invasion assays. h - j The formation of invadopodia in different groups of Hep3B and Huh7 cells was examined by immunofluorescence assays. k - m The function of invadopodia in different groups of Hep3B and Huh7 cells was examined by in situ zymography. A total of 150 cells per group were analyzed by two researchers in a blinded manner. Data are representative images or expressed as the mean +- SD of each group from three separate experiments. Bar scale in F = 100 mum. Bar scales in H and K = 20 mum

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8. Bin1 protein levels in muscles of R6/2 and control mice . (A-D) Western blots for Bin1 protein in the quadriceps femoris (A and B) and the gastrocnemius (C and D) of age-matched late-stage (10-13 wk of age) control (WT) and R6/2 mice. (A and C) Representative Western blots for Bin1 as well as beta-tubulin and GAPDH for normalization. Equal amounts of protein were loaded per lane (50 ug). (B and D) Bin1 band intensities normalized to beta-tubulin and GAPDH levels (relative to control) for the quadriceps femoris (B) and gastrocnemius (D). For A and B (quadriceps femoris), n = 4/group. For C and D (gastrocnemius), n = 6/group. *, Significantly different from control at P < 0.05. AU, arbitrary units.

PA5-16863

Antibody data