HA601275

GNAS Recombinant Mouse Monoclonal Antibody [A8F11-R]

cat.: HA601275

Product Type:	Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	A8F11-R

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 46 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within human GNAS aa 2-251.
Positive control:	NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, MCF7, human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue, rat pancreas tissue.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IF-Cell IHC-P	1:1,000 1:100 1:500-1:2,000
Uniprot #:	SwissProt: P63092 Human \| P84996 Human \| Q5JWF2 Human \| P63094 Mouse \| Q6R0H7 Mouse \| P63095 Rat \| Q63803 Rat
Alternative names:	Adenylate cyclase stimulating G alpha protein AHO Alternative gene product encoded by XL exon C20orf45 dJ309F20.1.1 dJ806M20.3.3 Extra large alphas protein GNAS GNAS complex locus GNAS1 GPSA Gs alpha subunit GSA GSP Guanine nucleotide binding protein (G protein) alpha stimulating activity polypeptide 1 Guanine nucleotide binding protein alpha stimulating activity polypeptide 1 Guanine nucleotide binding protein G(s) subunit alpha isoforms short Guanine nucleotide binding protein G(s) subunit alpha isoforms XLas Guanine nucleotide regulatory protein MGC33735 NESP NESP55 Neuroendocrine secretory protein PHP1A PHP1B PHP1C POH Protein ALEX Protein GNAS Protein NESP55 SCG6 Secretogranin VI XLalphas XLas

Images

Fig1: Western blot analysis of GNAS on different lysates with Mouse anti-GNAS antibody (HA601275) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: RAW264.7 cell lysate (20 µg/Lane)
Lane 3: C6 cell lysate (20 µg/Lane)
Lane 4: Human brain tissue lysate (40 µg/Lane)
Lane 5: Mouse brain tissue lysate (40 µg/Lane)
Lane 6: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 46 kDa
Observed band size: 44 kDa

Exposure time: 3 minutes;

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 (HA601275) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of MCF7 cells labeling GNAS with Mouse anti-GNAS antibody (HA601275) 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-GNAS antibody (HA601275) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Mouse anti-GNAS antibody (HA601275) at 1/500 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 (HA601275) at 1/500 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 mouse cerebellum tissue with Mouse anti-GNAS antibody (HA601275) at 1/500 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 (HA601275) at 1/500 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 rat cerebellum tissue with Mouse anti-GNAS antibody (HA601275) at 1/2,000 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 (HA601275) at 1/2,000 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 pancreas tissue with Mouse anti-GNAS antibody (HA601275) at 1/2,000 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 (HA601275) at 1/2,000 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.

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