HA500040

Glypican 5 Rabbit Polyclonal Antibody

cat.: HA500040

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:	1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 64 kDa.
Isotype:	IgG
Immunogen:	Synthetic peptide within human Glypican 5 aa 20-70.
Positive control:	HepG2 cell lysate, A549 cell lysate, Hela, rat kidney tissue, human lung tissue, mouse brain tissue, Hela.
Subcellular location:	Cell membrane, Membrane, Secreted.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:1,000 1:50-1:200 1:50-1:200 1:500-1:1,000
Uniprot #:	SwissProt: P78333 Human \| Q8CAL5 Mouse
Alternative names:	bA93M14.1 glypican 5 Glypican 5 precursor glypican proteoglycan 5 Glypican-5 GPC5 GPC5_HUMAN Secreted glypican-5

Images

Fig1: Western blot analysis of Glypican 5 on A549 (Human lung adenocarcinoma cell) cell lysate with Rabbit anti-Glypican 5 antibody (HA500040) at 1/1,000 dilution.

Lysates/proteins at 10 µg/Lane.
Exposure time: 110 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500040, 1/1,000 in 5% NFDM/TBST, overnight at 4°
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 64 kDa
Observed band size: 64 kDa

Fig2: Western blot analysis of Glypican 5 on Hep G2 (Human hepatocellular carcinoma cell) cell lysate with Rabbit anti-Glypican 5 antibody (HA500040) at 1/1,000 dilution.

Lysates/proteins at 10 µg/Lane.
Exposure time: 25 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500040, 1/1,000 in 5% NFDM/TBST, overnight at 4°
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 64 kDa
Observed band size: 64 kDa

	Fig3: ICC staining of Glypican 5 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA500040, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-Glypican 5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA500040, 1/100) for 30 minutes 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 human lung tissue using anti-Glypican 5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA500040, 1/100) for 30 minutes 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 mouse brain tissue using anti-Glypican 5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA500040, 1/100) for 30 minutes 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 Hela cells labeling Glypican 5.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500040, 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 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”.