beta IV Tubulin Recombinant Rabbit Monoclonal Antibody [PSH17-78]
cat.: HA751637
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH17-78
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 50 kDa
Isotype: IgG
Positive control: Jurkat cell lysate, SH-SY5Y cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, SH-SY5Y, Neuro-2a, C6, mouse brain tissue, mouse testis tissue, rat brain tissue, rat testis tissue.
Subcellular location: Cytoplasm, cytoskeleton.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000
1:250
1:5,000-1:20,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P04350 Human | Q9D6F9 Mouse
Entrez Gene: 29213 Rat
Alternative names: Beta 4 Beta 4 tubulin beta 5 beta four tubulin Dystonia 4 torsion (autosomal dominant) MC1R TBB4_HUMAN TUB B4 TUBB 4 tubb4 TUBB4A TUBB5 Tubulin 5 beta Tubulin beta 3 Tubulin beta 4 Tubulin beta 4 chain Tubulin beta 4A class IVa Tubulin beta 5 Tubulin beta IV Tubulin beta-4 chain
Images
HA751637_1.jpg Fig1: Western blot analysis of beta IV Tubulin on different lysates with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: SH-SY5Y cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (low expression) (20 µg/Lane)
Lane 4: PC-12 cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Mouse spleen tissue lysate (low expression) (40 µg/Lane)
Lane 8: Rat spleen tissue lysate (low expression) (40 µg/Lane)
Lane 9: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 50 kDa
Observed band size: 52 kDa

Exposure time: Lane 1-5: 25 seconds; Lane 6-9: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751637) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751637_2.jpg Fig2: Immunocytochemistry analysis of SH-SY5Y cells labeling beta IV Tubulin with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751637_3.jpg Fig3: Immunocytochemistry analysis of Neuro-2a cells labeling beta IV Tubulin with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751637_4.jpg Fig4: Immunocytochemistry analysis of C6 cells labeling beta IV Tubulin with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751637_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751637) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751637_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751637) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751637_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751637) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751637_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-beta IV Tubulin antibody (HA751637) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751637) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751637_9.jpg Fig9: Flow cytometric analysis of SH-SY5Y cells labeling beta IV Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751637, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA751637_10.jpg Fig10: Flow cytometric analysis of Neuro-2a cells labeling beta IV Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751637, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA751637_11.jpg Fig11: Flow cytometric analysis of C6 cells labeling beta IV Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751637, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA751637_12.jpg Fig12: beta IV Tubulin was immunoprecipitated from 0.2 mg Jurkat cell lysate with HA751637 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751637 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Jurkat cell lysate (input)
Lane 2: HA751637 IP in Jurkat cell lysate
Lane 3: Rabbit IgG instead of HA751637 in Jurkat cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 59 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.