HA722290

beta I Tubulin Recombinant Rabbit Monoclonal Antibody [JE01-23]

cat.: HA722290

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Monkey
Applications:	WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	JE01-23

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 50 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human beta I Tubulin aa 402-451 / 451.
Positive control:	HepG2 cell lysate, A549 cell lysate, K-562 cell lysate, PC-12 cell lysate, C6 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, HepG2, human breast cancer tissue, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Cytoplasm, cytoskeleton.
Recommended Dilutions: WB IF-Cell IHC-P FC IF-Tissue	1:1,000 1:250 3,000 1:1,000 1:200-1:500
Uniprot #:	SwissProt: Q9H4B7 Human \| A2AQ07 Mouse Entrez Gene: 679312 Rat
Alternative names:	2810484G07Rik beta 1 tubulin Beta tubulin 1, class VI Beta-tubulin Class VI beta tubulin dJ543J19.4 M(beta)1 TBB1_HUMAN TUBB1 Tubulin beta 1 class VI Tubulin beta-1 chain Tubulin, beta 1 tubulin, beta1

Images

Fig1: Western blot analysis of beta I Tubulin on different lysates with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: A549 cell lysate
Lane 3: K-562 cell lysate
Lane 4: PC-12 cell lysate
Lane 5: C6 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 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 (HA722290) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HepG2 cells labeling beta I Tubulin with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 I Tubulin antibody (HA722290) 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 (M1305-2, 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/3,000 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 (HA722290) at 1/3,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/3,000 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 (HA722290) at 1/3,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/3,000 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 (HA722290) at 1/3,000 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.

Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-beta I Tubulin antibody (HA722290) at 1/3,000 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 (HA722290) at 1/3,000 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.

Fig7: Flow cytometric analysis of HepG2 cells labeling beta I Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722290, 1μg/mL) (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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.