Type I keratin Antibody Sampler Kit Antibody Sampler Kit
cat.: HAK21150
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: P35527 Human | Q6RHW0 Mouse | Q8CIS9 Rat | P13645 Human | P02535 Mouse | Q6IFW6 Rat | Q99456 Human | Q64291 Mouse | Q6IFW5 Rat | P13646 Human | P08730 Mouse | P02533 Human | Q61781 Mouse | Q6IFV1 Rat | P19012 Human | Q61414 Mouse | Q6IFV3 Rat | P08779 Human | Q04695 Human | Q9QWL7 Mouse | Q6IFU8 Rat | P05783 Human | P05784 Mouse | Q5BJY9 Rat | P08727 Human | P19001 Mouse | Q63279 Rat | P35900 Human | P25030 Rat | Q01546 Human | Q7Z794 Human | P48668 Human | P13645 Human
Images
HAK21150_1.jpg Fig1: Western blot analysis of Keratin 12 / K12 on different lysates with Rabbit anti-Keratin 12 / K12 antibody (ET7111-24) at 1/1,000 dilution.

Lane 1: Mouse eyeball tissue lysate (40 µg/Lane)
Lane 2: C6 cell lysate (negative) (20 µg/Lane)
Lane 3: Rat eyeball tissue lysate (40 µg/Lane)

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

Exposure time: 3 minutes; 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 (ET7111-24) 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.
HAK21150_2.jpg Fig2: Western blot analysis of Cytokeratin 13 on different lysates with Rabbit anti-Cytokeratin 13 antibody (ET1611-55) at 1/2,000 dilution.

Lane 1: A431 cell lysate
Lane 2: HaCaT cell lysate
Lane 3: HEK-293 (-&low) cell lysate
Lane 4: Mouse lung tissue lysate
Lane 5: Mouse skin tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 50 kDa
Observed band size: 50 kDa

Exposure time: 10 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 (ET1611-55) 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.
HAK21150_3.jpg Fig3: Western blot analysis of Cytokeratin 14 on different lysates with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/2,000 dilution.

Lane 1: A431 cell lysate (15 µg/Lane)
Lane 2: PANC-1 cell lysate (negative) (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (negative) (15 µg/Lane)
Lane 4: PC-12 cell lysate (negative) (15 µg/Lane)
Lane 5: Mouse skin tissue lysate (20 µg/Lane)

Predicted band size: 51 kDa
Observed band size: 51 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 (ET1610-42) 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.
HAK21150_4.jpg Fig4: Western blot analysis of Cytokeratin 16 on different lysates with Rabbit anti-Cytokeratin 16 antibody (ET1610-17) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HaCaT cell lysate
Lane 3: HepG2 cell lysate
Lane 4: MCF7 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 51 kDa
Observed band size: 51 kDa

Exposure time: 16 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 (ET1610-17) 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.
HAK21150_5.jpg Fig5: Western blot analysis of Cytokeratin 17 on different lysates with Mouse anti-Cytokeratin 17 antibody (EM1901-29) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: MCF7 cell lysate (negative)
Lane 4: HepG2 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48 kDa
Observed band size: 48 kDa

Exposure time: 5 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 (EM1901-29) at 1/1,000 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.
HAK21150_6.jpg Fig6: Western blot analysis of Cytokeratin 18 on different lysates with Rabbit anti-Cytokeratin 18 antibody (ET1603-8) at 1/5,000 dilution.

Lane 1: A431 cell lysate
Lane 2: HeLa cell lysate
Lane 3: HT-29 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48 kDa
Observed band size: 48 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 (ET1603-8) 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.
HAK21150_7.jpg Fig7: Western blot analysis of pan Cytokeratin on different lysates with Mouse anti-pan Cytokeratin antibody (HA601138) at 1/2,000 dilution.

Lane 1: A431 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: MCF7 cell lysate (10 µg/Lane)
Lane 4: SK-Br-3 cell lysate (10 µg/Lane)
Lane 5: MDA-MB-231 cell lysate (10 µg/Lane)
Lane 6: MDA-MB-468 cell lysate (10 µg/Lane)
Lane 7: Human liver tissue lysate (20 µg/Lane)

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 (HA601138) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21150_8.jpg Fig8: Fluorescence multiplex immunohistochemical analysis of the human non-small cell lung cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD20 (HA721138, green), anti-CD68 (HA601115, gray), anti-PD-L1 (HA721176, cyan), anti-panCK (HA601138, magenta) and anti-CD3 (HA720082, yellow) on human non-small cell lung cancer. Panel B: anti- CD20 stained on B cells. Panel C: anti-CD68 stained on macrophage M1 and macrophage M2. Panel D: anti-PD-L1 stained on dendritic cells and macrophages cells. Panel E: anti-panCK stained on cancer cells. Panel F: anti-CD3 stained on T cells. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of HA721138 (1/1,500 dilution), HA601115 (1/2,000 dilution), HA721176 (1/1,000 dilution), HA601138 (1/3,000 dilution), and HA720082 (1/500 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HAK21150_9.jpg Fig9: Immunofluorescence analysis of paraffin-embedded human stomach cancer tissue labeling pan Cytokeratin with Mouse anti-pan Cytokeratin antibody (HA601138) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA601138, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21150_10.jpg Fig10: Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 9 (ET1612-77) and Vimentin (EM0401).

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 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 9 (ET1612-77, red) at 1/50 dilution and Vimentin (EM0401, green) at 1/500 dilution at +4℃ overnight, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) and Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) were used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21150_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue with Rabbit anti-Cytokeratin 10 antibody (ET1609-75) 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 (ET1609-75) 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.
HAK21150_12.jpg Fig12: Immunocytochemistry analysis of A431 (positive) and HEK-293 (negative) labeling Cytokeratin 13 with Rabbit anti-Cytokeratin 13 antibody (ET1611-55) at 1/500 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-Cytokeratin 13 antibody (ET1611-55) 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.
HAK21150_13.jpg Fig13: Flow cytometric analysis of HEK-293 (left, negative) and A431 (right, positive) cells labeling Cytokeratin 13.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-55, 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).
HAK21150_14.jpg Fig14: Immunocytochemistry analysis of A431 cells labeling Cytokeratin 14 with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/500 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-Cytokeratin 14 antibody (ET1610-42) 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.
HAK21150_15.jpg Fig15: Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 16 with Rabbit anti-Cytokeratin 16 antibody (ET1610-17) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-17, red) at 1/200 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
HAK21150_16.jpg Fig16: Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-AQP1 (ET1703-34, Violet), anti-CK18 (ET1603-8, Green) and anti-Calbindin (ET1702-54, Yellow) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1703-34 (1/5,000 dilution), ET1603-8 (1/3,000 dilution) and ET1702-54 (1/4,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
HAK21150_17.jpg Fig17: Fluorescence multiplex immunohistochemical analysis of mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Red), anti-Glucagon (ET1702-20, Green), anti-Insulin (ET1601-12, White), anti-CK19 (ET1601-6, Magenta) and anti-aSMA (ET1607-53, Yellow) on mouse pancreas. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1702-20 (1/6,000 dilution), ET1601-12 (1/8,000 dilution), ET1601-6 (1/5,000 dilution) and ET1607-53 (1/10,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HAK21150_18.jpg Fig18: Immunocytochemistry analysis of MCF-7 cells labeling Cytokeratin 19 with Rabbit anti-Cytokeratin 19 antibody (ET1601-6) at 1/500 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-Cytokeratin 19 antibody (ET1601-6) 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.
HAK21150_19.jpg Fig19: Cytokeratin 20 was immunoprecipitated from 0.2 mg HT-29 cell lysate with ET1601-8 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-8 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: HT-29 cell lysate (input)
Lane 2: ET1601-8 IP in HT-29 cell lysate
Lane 3: Rabbit IgG instead of ET1601-8 in HT-29 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.