Lysosomal Marker Antibody Sampler Kit
cat.: HAK21188
Product Type: Antibody Sampler Kit
Species reactivity: Human
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.
Concentration: 1ug/ul
Uniprot #: SwissProt: P11279 Human | P11438 Mouse | P14562 Rat | P13473 Human | P17047 Mouse | P17046 Rat | P07858 Human | P07339 Human | P18242 Mouse | P24268 Rat | Q9GZU1 Human | Q99J21 Mouse | P08962 Human | P04062 Human | P17439 Mouse | Q14108 Human | O35114 Mouse | P27615 Rat | P38606 Human | P50516 Mouse
Entrez Gene: 684536 Rat | 685232 Rat
Images
HAK21188_1.jpg Fig1: Western blot analysis of LAMP1 on different lysates with Rabbit anti-LAMP1 antibody (HA722827) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: A431 cell lysate
Lane 4: MCF7 cell lysate
Lane 5: MDA-MB-231 cell lysate
Lane 6: HeLa cell lysate
Lane 7: HeLa cell lysate treated with deglycosylation
Lane 8: NIH/3T3 cell lysate
Lane 9: NIH/3T3 cell lysate treated with deglycosylation

Lysates/proteins at 20 µg/Lane.

Predicted band size: 44 kDa
Observed band size: 120/44 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 (HA722827) 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.
HAK21188_2.jpg Fig2: Application: IHC-Fr

Species: Mouse

Site: Kidney

Sample: Frozen section

Antibody concentration: 1/1,000

Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.
HAK21188_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-LAMP1 antibody (HA722827) 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 (HA722827) 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.
HAK21188_4.jpg Fig4: LAMP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722827 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722827 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA722827 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722827 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801
HAK21188_5.jpg Fig5: Western blot analysis of LAMP2a on different lysates with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/5,000 dilution.

Lane 1: SK-MEL-28 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: JAR cell lysate (20 µg/Lane)
Lane 4: U-937 cell lysate (20 µg/Lane)
Lane 5: RAW264.7 cell lysate (15 µg/Lane)
Lane 6: NIH/3T3 cell lysate (15 µg/Lane)
Lane 7: PC-12 cell lysate (15 µg/Lane)
Lane 8: Mouse liver tissue lysate (30 µg/Lane)
Lane 9: Rat liver tissue lysate (30 µg/Lane)
Lane 10: Rat lung tissue lysate (30 µg/Lane)

Predicted band size: 45 kDa
Observed band size: 70-140 kDa

Exposure time: Lane 1-4: 2 minutes 37 seconds; Lane 5-10: 5 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 (ET1601-24) 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.
HAK21188_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-LAMP2a antibody (ET1601-24) at 1/500 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 (ET1601-24) at 1/500 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.
HAK21188_7.jpg Fig7: LAMP2a was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with ET1601-24 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-24 at 1/2,000 dilution. HRP Conjugated Rabbit IgG kappa light chain antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: SK-MEL-28 cell lysate (Input)
Lane 2: ET1601-24 IP in SK-MEL-28 cell lysate
Lane 3: Rabbit IgG instead of ET1601-24 in SK-MEL-28 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 20 seconds; ECL: K1801
HAK21188_8.jpg Fig8: Western blot analysis of Cathepsin D on different lysates with Rabbit anti-Cathepsin D antibody (ET1608-49) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane)
Lane 3: Neuro-2a cell lysate (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)

Predicted band size: 45/28 kDa
Observed band size: 45/28 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 (ET1608-49) 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.
HAK21188_9.jpg Fig9: Western blot analysis of Mucolipin-1 on different lysates with Rabbit anti-Mucolipin-1 antibody (HA723054) at 1/2,000 dilution.

Lane 1: SK-MEL-28 cell lysate
Lane 2: NCI-H226 cell lysate
Lane 3: Ramos cell lysate (negative)
Lane 4: TT cell lysate
Lane 5: Mouse brain tissue lysate
Lane 6: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 65 kDa
Observed band size: 35-40 kDa (PMID: 16257972)

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 (HA723054) 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.
HAK21188_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Mucolipin-1 antibody (HA723054) 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 (HA723054) 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.
HAK21188_11.jpg Fig11: Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (HA722731) at 1/1,000 dilution.

Lane 1: SK-MEL-28 cell lysate
Lane 2: Jurkat cell lysate (negative)
Lane 3: U-87 MG cell lysate
Lane 4: HUVEC cell lysate
Lane 5: K-562 cell lysate
Lane 6: HEK-293 cell lysate
Lane 7: HL-60 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 26 kDa
Observed band size: 30-65 kDa

Exposure time: Lane 1: 4 seconds; Lane 2-7: 18 seconds; ECL: K1802;

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 (HA722731) 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.
HAK21188_12.jpg Fig12: Flow cytometric analysis of K-562 cells labeling CD63.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722731, 1μg/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).
HAK21188_13.jpg Fig13: Western blot analysis of LIMPII on different lysates with Rabbit anti-LIMPII antibody (HA721967) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HepG2 cell lysate (15 µg/Lane)
Lane 3: SH-SY5Y cell lysate (15 µg/Lane)
Lane 4: Neuro-2a cell lysate (15 µg/Lane)
Lane 5: MEF cell lysate (15 µg/Lane)
Lane 6: PC-12 cell lysate (15 µg/Lane)
Lane 7: COS-1 cell lysate (15 µg/Lane)
Lane 8: Mouse kidney tissue lysate (30 µg/Lane)
Lane 9: Rat kidney tissue lysate (30 µg/Lane)
Lane 10: Mouse liver tissue lysate (30 µg/Lane)
Lane 11: Rat liver tissue lysate (30 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 80 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 (HA721967) at 1/5,000 dilution and competitor's antibody at 1/5,000 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.
HAK21188_14.jpg Fig14: Immunocytochemistry analysis of Neuro-2a cells labeling LIMPII with Rabbit anti-LIMPII antibody (HA721967) 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-LIMPII antibody (HA721967) 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.
HAK21188_15.jpg Fig15: Western blot analysis of ATP6V1A on different lysates with Rabbit anti-ATP6V1A antibody (HA724200) at 1/20,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: COS-1 cell lysate
Lane 5: Mouse kidney tissue lysate
Lane 6: Rat kidney tissue lysate
Lane 7: Mouse brain tissue lysate
Lane 8: Rat brain tissue lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 68 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 (HA724200) at 1/20,000 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.
HAK21188_16.jpg Fig16: Immunocytochemistry analysis of HeLa cells labeling ATP6V1A with Rabbit anti-ATP6V1A antibody (HA724200) 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-ATP6V1A antibody (HA724200) 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 (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.
HAK21188_17.jpg Fig17: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP6V1A antibody (HA724200) at 1/8,000 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 (HA724200) at 1/8,000 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.
HAK21188_18.jpg Fig18: ATP6V1A was immunoprecipitated from 0.2 mg 293T cell lysate with HA724200 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724200 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: 293T cell lysate (input)
Lane 2: HA724200 IP in 293T cell lysate
Lane 3: Rabbit IgG instead of HA724200 in 293T cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 10 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.