M1305-2

beta Tubulin Mouse Monoclonal Antibody [A1-A4]

cat.: M1305-2

Product Type:	Mouse monoclonal IgG3, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	A1-A4

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 50 kDa
Isotype:	IgG3
Immunogen:	Synthetic peptide within Human Beta tubulin aa 151-200 / 444.
Positive control:	Hela cell lysates, NIH/3T3 cell lysates, PC-12 cell lysates, SKOV-3 cells, HeLa, 293, human tonsil tissue, human spleen tissue, human colon tissue, rat brain tissue, NIH/3T3.
Subcellular location:	Cytoplasm, Cytoskeleton, Microtubule.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000-1:20,000 1:100-1:500 1:50-1:200 1:100-1:1,000
Uniprot #:	SwissProt: P07437 Human \| P99024 Mouse \| P69897 Rat
Alternative names:	beta 3 tubulin beta-4 CDCBM CDCBM1 CFEOM3 CFEOM3A FEOM3 M(beta)3 M(beta)6 MC1R Neuron specific beta III Tubulin Neuron-specific class III beta-tubulin QccE-11995 QccE-15186 TBB3_HUMAN Tubb 3 TUBB3 TUBB4 Tubulin beta 3 Tubulin beta 3 chain Tubulin beta 4 Tubulin beta III Tubulin beta-3 chain Tubulin beta-4 chain Tubulin beta-III

Images

Fig1: Western blot analysis of beta Tubulin on Hela cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

Hela cell lysates at 10 µg/Lane.

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

8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of beta Tubulin on NIH/3T3 cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

NIH/3T3 cell lysates at 10 µg/Lane.

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

8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of beta Tubulin on PC-12 cell lysates with Mouse anti-beta Tubulin antibody (M1305-2).

PC-12 cell lysates at 10 µg/Lane.

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

8% 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 (M1305-2) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of beta Tubulin on different lysates with Rabbit anti-beta Tubulin antibody (M1305-2) at 1/1,000 dilution.

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

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 2 seconds;

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 (M1305-2) 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.

	Fig5: Immunocytochemistry analysis of HeLa cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 dilution. Cells were fixed in 100% precooled methanol 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 Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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.
	Fig6: Immunocytochemistry analysis of 293 cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 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 Mouse anti-beta Tubulin antibody (M1305-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.

	Fig8: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.
	Fig9: Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.
	Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-beta tubulin antibody. Counter stained with hematoxylin.
	Fig11: Flow cytometric analysis of HeLa cells labeling beta Tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

	Fig12: Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig13: Immunocytochemistry analysis of PC-12 cells labeling beta Tubulin with Rabbit anti-beta Tubulin antibody (M1305-2) at 1/100 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 Tubulin antibody (M1305-2) at 1/100 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.
	Fig14: Flow cytometric analysis of HeLa cells labeling beta Tubulin. Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 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).

Fig15: Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 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).

Fig16: Flow cytometric analysis of PC-12 HeLa cells labeling beta Tubulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (M1305-2, 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).

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