Gluconeogenesis Antibody Sampler Kit
cat.: HAK21135
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB
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.
Uniprot #: SwissProt: P09104 Human | P17183 Mouse | P07323 Rat | P18669 Human | Q9DBJ1 Mouse | P25113 Rat | P06744 Human | P06733 Human | P17182 Mouse | P04764 Rat | P00558 Human | P09411 Mouse | P16617 Rat | Q16822 Human | Q8BH04 Mouse | P09467 Human | Q9QXD6 Mouse | P19112 Rat | P11498 Human | Q05920 Mouse | P52873 Rat | P35558 Human
Entrez Gene: 361042 Rat
Images
HAK21135_1.jpg Fig1: Western blot analysis of PGK1 on different lysates with Rabbit anti-PGK1 antibody (ET1609-63) at 1/2,000 dilution.

Lane 1: HT-29 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C2C12 cell lysate
Lane 4: C6 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 45 kDa

Exposure time: 9 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 (ET1609-63) at 1/2,000 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.
HAK21135_2.jpg Fig2: Western blot analysis of Glucose 6 phosphate isomerase on different lysates with Rabbit anti-Glucose 6 phosphate isomerase antibody (ET7108-01) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Glucose 6 phosphate isomerase KD cell lysate

Lysates/proteins at 10 µg/Lane.
Predicted band size: 63 kDa
Observed band size: 54 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 (ET7108-01) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21135_3.jpg Fig3: Western blot analysis of ENO1 on different lysates with Rabbit anti-ENO1 antibody (ET1705-56) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
Lane 4: C6 cell lysate (20 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: Rat brain tissue lysate (20 µg/Lane)
Lane 7: Zebrafish tissue lysate (20 µg/Lane)

Predicted band size: 47 kDa
Observed band size: 47 kDa

Exposure time: 6 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 (ET1705-56) at 1/2,000 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.
HAK21135_4.jpg Fig4: Western blot analysis of PCK2 on different lysates with Rabbit anti-PCK2 antibody (ET7107-29) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-PCK2 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 71 kDa
Observed band size: 67 kDa

Exposure time: 21 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 (ET7107-29) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21135_5.jpg Fig5: Western blot analysis of PCB on different lysates with Rabbit anti-PCB antibody (HA721698) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: MCF7 cell lysate
Lane 3: A549 cell lysate
Lane 4: Huh7 cell lysate
Lane 5: Hela cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: Rat brain tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 130 kDa

Exposure time: 24 seconds;

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 (HA721698) 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 1/50,000 dilution was used for 1 hour at room temperature.
HAK21135_6.jpg Fig6: Western blot analysis of PCB on different lysates with Rabbit anti-PCB antibody (HA721698) at 1/2,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-PCB KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 130 kDa

Exposure time: 1 minute; 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 (HA721698) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21135_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-NSE antibody (ET1610-96) 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 (ET1610-96) 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.
HAK21135_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-FBP1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET7109-82, 1/50) 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.
HAK21135_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-PCB antibody (HA721698) 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 (HA721698) 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.
HAK21135_10.jpg Fig10: Immunocytochemistry analysis of MCF7 cells labeling PCK2 with Rabbit anti-PCK2 antibody (ET7107-29) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-PCK2 antibody (ET7107-29) at 1/100 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
HAK21135_11.jpg Fig11: Immunocytochemistry analysis of Neuro-2a cells labeling PCK2 with Rabbit anti-PCK2 antibody (ET7107-29) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-PCK2 antibody (ET7107-29) at 1/100 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
HAK21135_12.jpg Fig12: Immunocytochemistry analysis of HepG2 cells labeling PCB with Rabbit anti-PCB antibody (HA721698) 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 Rabbit anti-PCB antibody (HA721698) at 1/100 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.