Glycolysis Antibody Sampler Kit II
cat.: HAK21101
Product Type: Antibody Sampler Kit
Species reactivity: Human, Mouse
Applications: WB, IHC-P
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: P04075 Human | P05064 Mouse | P05065 Rat | P06733 Human | P17182 Mouse | P04764 Rat | P09104 Human | P17183 Mouse | P07323 Rat | Q15118 Human | Q8BFP9 Mouse | Q63065 Rat | O60825 Human | P70265 Mouse | Q16875 Human | O35552 Rat | Q3U3S6 Mouse | P18669 Human | Q9DBJ1 Mouse | P25113 Rat
Images
HAK21101_1.jpg Fig1: Western blot analysis of Aldolase on different lysates with Rabbit anti-Aldolase antibody (ET1705-91) at 1/1,000 dilution.

Lane 1: A549 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: Mouse liver tissue lysate (20 µg/Lane)
Lane 4: Rat liver tissue lysate (20 µg/Lane)
Lane 5: Rat spleen tissue lysate (20 µg/Lane)

Predicted band size: 39 kDa
Observed band size: 39 kDa

Exposure time: 1 minute 20 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 (ET1705-91) 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 1:100,000 dilution was used for 1 hour at room temperature.
HAK21101_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Aldolase antibody (ET1705-91) 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 (ET1705-91) 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.
HAK21101_3.jpg Fig3: Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD PDK1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 46 kDa

Exposure time: 3 minutes; 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 (ET1704-66) at 1/1,000 dilution was used in primary antibody diluent at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21101_4.jpg Fig4: PDK1 was immunoprecipitated from 0.2 mg mouse heart tissue lysate with ET1704-66 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1704-66 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Mouse heart tissue lysate (input)
Lane 2: ET1704-66 IP in mouse heart tissue lysate
Lane 3: Rabbit IgG instead of ET1704-66 in mouse heart tissue lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
HAK21101_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) 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 (ET1704-66) 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.
HAK21101_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Aldolase antibody (ET1705-91) 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 (ET1705-91) 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.
HAK21101_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-PFKFB2 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 (HA500200, 1/400) 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.
HAK21101_8.jpg Fig8: Western blot analysis of NSE on different lysates with Rabbit anti-NSE antibody (ET1610-96) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: HEK-293 cell lysate (10 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (10 µg/Lane)
Lane 6: SH-SY5Y cell lysate (10 µg/Lane)

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

Exposure time: 1 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 (ET1610-96) 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 1:100,000 dilution was used for 1 hour at room temperature.
HAK21101_9.jpg Fig9: All lanes: Western blot analysis of PFKFB3 with anti-PFKFB3 antibody [JM43-43] (ET1705-66) at 1/500 dilution.

Lane 1: Wild-type Hela whole cell lysate.
Lane 2: PFKFB3 knockout Hela whole cell lysate.

ET1705-66 was shown to specifically react with PFKFB3 in wild-type Hela cells. No band was observed when PFKFB3 knockout sample was tested. Wild-type and PFKFB3 knockout 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 Anti-PFKFB3 antibody (ET1705-66, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China).
HAK21101_10.jpg Fig10: Western blot analysis of PGAM1 on different lysates using anti-PGAM1 antibody at 1/2,000 dilution.
Positive control:
Lane 1: A431
Lane 2: A549
Lane 3: Rat brain
Lane 4: Mouse brain
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.