Mitoxyperilysis Antibody Sampler Kit
cat.: HAK21205
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.
Concentration: 1ug/ul
Uniprot #: SwissProt: O60603 Human | Q9QUN7 Mouse | Q07812 Human | Q07813 Mouse | Q63690 Rat | Q16611 Human | P55957 Human | P70444 Mouse | Q96P20 Human | Q8R4B8 Mouse | P01584 Human | P10749 Mouse | Q63264 Rat | Q6R327 Human | P31749 Human | P31751 Human | Q9Y243 Human | P31750 Mouse | P47196 Rat
Entrez Gene: 287362 Rat
Images
HAK21205_1.jpg Fig1: Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (ET1603-34) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: bEnd.3 cell lysate
Lane 5: C2C12 cell lysate
Lane 6: PC-12 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 21 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 (ET1603-34) at 1/20,000 dilution and competitor's antibody at 1/1,000 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.
HAK21205_2.jpg Fig2: All lanes: Western blot analysis of Bax with anti-Bax antibody [SZ3-07] (ET1603-34) at 1/1,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (20 µg).
Lane 3/4: Bax knockout Hela whole cell lysate (20 µg).

ET1603-34 was shown to specifically react with Bax in wild-type Hela cells. No band was observed when Bax knockout samples were tested. Wild-type and Bax 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 antibody (ET1603-34, 1/1,000) and Loading control antibody (Rabbit anti-β-actin, R1207-1, 1/1,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
HAK21205_3.jpg Fig3: Immunocytochemistry analysis of C2C12 cells labeling BAX with Rabbit anti-BAX antibody (ET1603-34) 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-BAX antibody (ET1603-34) 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.
HAK21205_4.jpg Fig4: All lanes: Western blot analysis of TLR2 with anti-TLR2 antibody [JM22-41] (ET1705-92) at 1/500 dilution.

Lane 1: Wild-type THP-1 whole cell lysate.
Lane 2: TLR2 knockout THP-1 whole cell lysate.

ET1705-92 was shown to specifically react with TLR2 in wild-type THP-1 cells. No band was observed when TLR2 knockout samples were tested. Wild-type and TLR2 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-TLR2 antibody (ET1705-92, 1/500) and Anti-β-actin antibody (R1207-1, 1/1,000) 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).
HAK21205_5.jpg Fig5: Western blot analysis of Bak on different lysates with Rabbit anti-Bak antibody (ET1608-21) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: AGS cell lysate
Lane 3: HEK-293 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 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 (ET1608-21) 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.
HAK21205_6.jpg Fig6: Western blot analysis of Bak on different lysates with Rabbit anti-Bak antibody (ET1608-21) at 1/2,000 dilution.

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

Lysates/proteins at 10 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Exposure time: 120 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 (ET1608-21) at 1/2,000 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.
HAK21205_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Bak antibody (ET1608-21) 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 (ET1608-21) 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.
HAK21205_8.jpg Fig8: Western blot analysis of Bid on different lysates with Rabbit anti-Bid antibody (HA723593) at 1/5,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: HT-29 cell lysate
Lane 3: A549 cell lysate
Lane 4: RAW264.7 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 22 kDa
Observed band size: 22 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 (HA723593) at 1/5,000 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.
HAK21205_9.jpg Fig9: Immunocytochemistry analysis of RAW264.7 cells treated with 10μg/mL LPS for 8 hours labeling NLRP3 with Rabbit anti-NLRP3 antibody (ET1610-93) 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-NLRP3 antibody (ET1610-93) 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 (HA601187, 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.
HAK21205_10.jpg Fig10: Western blot analysis of IL-1 beta on different lysates with Mouse anti-IL-1 beta antibody (HA601036) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 80nM TPA overnight then replaced with 100ng/mL LPS for 6 hours add 300ng/mL BFA for last 3 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 35/17 kDa

Exposure time: 4 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 (HA601036) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21205_11.jpg Fig11: Application: IF-Tissue

Species: Human

Site: tonsil

Sample: Paraffin-embedded section

Antibody concentration: 1/200
HAK21205_12.jpg Fig12: Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (ET1607-73) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: MCF7 treated with 100nM Calyculin A for 30 minutes cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 100ng/mL PDGF for 1 hour cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 53 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 (ET1607-73) at 1/5,000 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.
HAK21205_13.jpg Fig13: Immunocytochemistry analysis of HeLa cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT (S473) with Rabbit anti-Phospho-AKT (S473) antibody (ET1607-73) at 1/200 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-Phospho-AKT (S473) antibody (ET1607-73) 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.
HAK21205_14.jpg Fig14: Immunocytochemistry analysis of NIH/3T3 cells treated with or without 100ng/mL PDGF for 1 hour labeling Phospho-AKT (S473) with Rabbit anti-Phospho-AKT (S473) antibody (ET1607-73) 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-Phospho-AKT (S473) antibody (ET1607-73) 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.
HAK21205_15.jpg Fig15: Application: IF-tissue

Species: Mouse

Site: Lung

Sample: Paraffin-embedded section

Antibody concentration: 1/200
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.