HA500372

MLST8 Rabbit Polyclonal Antibody

cat.: HA500372

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 36 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human MLST8 aa 1-250.
Positive control:	C2C12 cell lysate, L6 cell lysate, mouse brain tissue lysate, human brain tissue lysate, rat brain tissue lysate, 293 cell lysate, MCF-7 cell lysate, PC-3M, rat heart tissue, mouse skeletal muscle tissue, PC-3.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:1,000 1:200 1:800 1:1,000
Uniprot #:	SwissProt: Q9BVC4 Human \| Q9DCJ1 Mouse \| Q9Z2K5 Rat
Alternative names:	G protein beta subunit like G protein beta subunit-like Gable GbetaL GBL GBL protein LST8 LST8_HUMAN Mammalian lethal with SEC13 protein 8 MGC111011 mLST8 MTOR associated protein LST8 homolog (S. cerevisiae) POP3 POP3 homolog (S. pombe) Protein GbetaL Target of rapamycin complex subunit LST8 TORC subunit LST8 WAT1 WAT1 homolog (S. pombe)

Images

Fig1: Western blot analysis of MLST8 on different lysates with Rabbit anti-MLST8 antibody (HA500372) at 1/1,000 dilution.

Lane 1: C2C12 cell lysate (10 µg/Lane)
Lane 2: L6 cell lysate (10 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Human brain tissue lysate (20 µg/Lane)
Lane 5: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 40 kDa
Observed band size: 36 kDa

Exposure time: 2 minutes;

12% 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 (HA500372) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of MLST8 on different lysates with Rabbit anti-MLST8 antibody (HA500372) at 1/500 dilution.

Lane 1: 293 cell lysate
Lane 2: MCF-7 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 40 kDa
Observed band size: 36 kDa

Exposure time: 2 minutes;

12% 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 (HA500372) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of PC-3M cells labeling MLST8 with Rabbit anti-MLST8 antibody (HA500372) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-MLST8 antibody (HA500372) at 1/200 dilution in 2% negative goat serum 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-MLST8 antibody (HA500372) at 1/800 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 (HA500372) at 1/800 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 skeletal muscle tissue with Rabbit anti-MLST8 antibody (HA500372) at 1/800 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 (HA500372) at 1/800 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: Flow cytometric analysis of PC-3 cells labeling MLST8.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500372, 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).

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