HA721314

G3BP Recombinant Rabbit Monoclonal Antibody [JE35-20]

cat.: HA721314

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	JE35-20

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: 52 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human G3BP aa 367-466 / 466.
Positive control:	A549 cell lysate, LoVo cell lysate, Jurkat cell lysate, Neuro-2a cell lysate, C6 cell lysate, human colon tissue, mouse brain tissue, rat cerebellum tissue, A549, LoVo, Neuro-2a.
Subcellular location:	Cytoplasm, Nucleus
Recommended Dilutions: WB IHC-P IF-Cell FC	1:1,000 1:200-1:1,000 1:50 1:1,000
Uniprot #:	SwissProt: Q13283 Human \| P97855 mouse \| P50904 rat
Alternative names:	ATP dependent DNA helicase VIII ATP-dependent DNA helicase VIII G3BP G3BP stress granule assembly factor 1 G3BP-1 G3bp1 G3BP1_HUMAN GAP binding protein GAP SH3 domain binding protein 1 GAP SH3 domain-binding protein 1 GTPase activating protein (SH3 domain) binding protein 1 hDH VIII Human DNA helicase VIII MGC111040 Ras GTPase activating protein binding protein 1 Ras GTPase activating protein SH3 domain binding protein Ras GTPase-activating protein-binding protein 1 RasGAP associated endoribonuclease G3BP

Images

Fig1: Western blot analysis of G3BP on different lysates with Rabbit anti-G3BP antibody (HA721314) at 1/1,000 dilution.

Lane 1: A549 cell lysate
Lane 2: LoVo cell lysate
Lane 3: Jurkat cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 52 kDa
Observed band size: 60 kDa

Exposure time: 20 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 (HA721314) 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: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-G3BP antibody (HA721314) at 1/200 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 (HA721314) at 1/200 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-G3BP antibody (HA721314) at 1/200 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 (HA721314) at 1/200 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 rat cerebellum tissue with Rabbit anti-G3BP antibody (HA721314) at 1/1,000 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 (HA721314) at 1/1,000 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: Flow cytometric analysis of A549 cells labeling G3BP. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721314, 1ug/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 Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig6: Immunocytochemistry analysis of LoVo cells labeling G3BP with Rabbit anti-G3BP antibody (HA721314) at 1/50 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-G3BP antibody (HA721314) at 1/50 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.

Fig7: Immunocytochemistry analysis of Neuro-2a cells labeling G3BP with Rabbit anti-G3BP antibody (HA721314) at 1/50 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-G3BP antibody (HA721314) at 1/50 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.

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