ET1601-10

Glucose Transporter GLUT1 Recombinant Rabbit Monoclonal Antibody [SA0377]

cat.: ET1601-10

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	SA0377

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 54 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human GLUT1 aa 443-492 / 492.
Positive control:	HeLa cell lysate, PC-12 cell lysate, HeLa, HT-29 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, L-929 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, Jurkat, NIH/3T3, C6, human liver tissue, human placenta tissue, human liver carcinoma tissue, human kidney tissue, mouse liver tissue, mouse kidney tissue, HepG2, human lung cancer tissue, human liver tissue.
Subcellular location:	Cell membrane, Melanosome
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue FC	1:50,000-1:100,000 1:500-1:1,000 1:5,000-1:10,000 1:500-1:1,000 1:500-1:1,000
Uniprot #:	SwissProt: P11166 Human \| P17809 Mouse \| P11167 Rat
Alternative names:	Choreoathetosis/spasticity episodic (paroxysmal choreoathetosis/spasticity) CSE DYT17 DYT18 DYT9 EIG12 erythrocyte/brain Erythrocyte/hepatoma glucose transporter facilitated glucose transporter member 1 Glucose transporter 1 Glucose transporter type 1 Glucose transporter type 1, erythrocyte/brain GLUT GLUT-1 GLUT1 GLUT1DS GLUTB GT1 GTG1 Gtg3 GTR1_HUMAN HepG2 glucose transporter HTLVR Human T cell leukemia virus (I and II) receptor MGC141895 MGC141896 PED RATGTG1 Receptor for HTLV 1 and HTLV 2 SLC2A1 Solute carrier family 2 (facilitated glucose transporter), member 1 Solute carrier family 2 Solute carrier family 2, facilitated glucose transporter member 1

Images

Fig1: Western blot analysis of Glucose Transporter GLUT1 on different lysates with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/50,000 dilution and competitor's antibody at 1/50,000 dilution.

Lane 1: HeLa cell lysate (no heat) (20 µg/Lane)
Lane 2: HT-29 cell lysate (no heat) (20 µg/Lane)
Lane 3: HepG2 cell lysate (no heat) (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (no heat) (20 µg/Lane)
Lane 5: L-929 cell lysate (no heat) (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (no heat) (20 µg/Lane)
Lane 7: Rat brain tissue lysate (no heat) (20 µg/Lane)

Notice: no heat means the lysate is not boiled.

Predicted band size: 54 kDa
Observed band size: 45-60 kDa

Exposure time: Lane 1-7 (left): 20 seconds; Lane 1-7 (right): 1 minute 30 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 (ET1601-10) at 1/50,000 dilution and competitor's antibody at 1/50,000 dilution were 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: Western blot analysis of Glucose Transporter GLUT1 on different lysates with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/50,000 dilution.

Lane 1: HeLa cell lysate (RIPA lysis) (10 µg/Lane)
Lane 2: HeLa cell lysate (hot lysis) (10 µg/Lane)
Lane 3: PC-12 cell lysate (RIPA lysis) (10 µg/Lane)
Lane 4: PC-12 cell lysate (hot lysis) (10 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 30 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 (ET1601-10) at 1/50,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of GLUT1 on different lysates with Rabbit anti-GLUT1 antibody (ET1601-10) at 1/50,000 dilution.

Lane 1: Hela-si NT cell lysate (no heat)
Lane 2: Hela-si GLUT1#1 cell lysate (no heat)
Lane 3: Hela-si GLUT1#2 cell lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 45-60 kDa

Exposure time: 1minute 50 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

ET1601-10 was shown to specifically react with GLUT1 in Hela-si NT cells. Weakened band was observed when Hela-si GLUT1 sample was tested. Hela-si NT and Hela-si GLUT1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1601-10, 1/50,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig4: Immunocytochemistry analysis of HeLa cells labeling Glucose Transporter GLUT1 with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 dilution and competitor's antibody at 1/200 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 Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 dilution and competitor's antibody at 1/200 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.

	Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling Glucose Transporter GLUT1 with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 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 Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 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.
	Fig6: Flow cytometric analysis of Jurkat cells labeling Glucose Transporter GLUT1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-10, red) at 1/1,000 dilution and competitor's antibody (red) at 1/50 dilution, compared with Rabbit IgG Isotype Control (blue). 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).
	Fig7: Immunocytochemistry analysis of C6 cells labeling Glucose Transporter GLUT1 with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 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 Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/500 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.

	Fig8: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) at 1/5,000 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) at 1/5,000 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/10,000 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 (ET1601-10) at 1/10,000 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.
	Fig11: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) at 1/5,000 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.

	Fig12: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) at 1/5,000 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.
	Fig13: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) 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.
	Fig14: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Glucose Transporter GLUT1 antibody (ET1601-10) at 1/5,000 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 (ET1601-10) 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.
	Fig15: Application: IF-tissue Species: Human Site: Liver Sample: Paraffin-embedded section Antibody concentration: 1/500 Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Fig16: Application: IF-tissue

Species: Human

Site: Placenta

Sample: Paraffin-embedded section

Antibody concentration: 1/500

Fig17: Flow cytometric analysis of HepG2 cells labeling Glucose Transporter GLUT1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-10, 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).

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