Cytokeratin 5+6 Recombinant Rabbit Monoclonal Antibody [PSH0-68]
cat.: HA721456
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH0-68
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 60 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Cytokeratin 6 186-235.
Positive control: A431 cell lysate, NCI-H226 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HeLa, A431.
Subcellular location: Cytoskeleton.
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:1,000
1:100
1:500-1:1,000
Uniprot #: SwissProt: P02538 Human | P04259 Human | P13647 Human
Alternative names: 58 kDa cytokeratin CK-5 CK5 Cytokeratin-5 Cytokeratin5 DDD DDD1 EBS2 epidermolysis bullosa simplex 2 Dowling-Meara/Kobner/Weber-Cockayne types K2C5_HUMAN K5 keratin 5 (epidermolysis bullosa simplex, Dowling-Meara/Kobner/Weber-Cockayne types) Keratin 5 Keratin keratin complex 2, basic, gene 5 keratin, type II cytoskeletal 5 Keratin-5 Keratin5 KRT 5 Krt5 KRT5A type II cytoskeletal 5 Type-II keratin Kb5 CK6 CK 6A CK 6B CK 6C CK 6D CK 6E CK-6B CK-6C CK-6E Cytokeratin 6a Cytokeratin 6B Cytokeratin 6C Cytokeratin 6D Cytokeratin 6E Cytokeratin-6B Cytokeratin-6C Cytokeratin-6E K2C6C_HUMAN K6a keratin K6b keratin K6C K6c keratin K6d keratin K6e keratin Keratin Keratin K6h Keratin type II cytoskeletal 6A Keratin type II cytoskeletal 6B Keratin type II cytoskeletal 6C Keratin type II cytoskeletal 6D Keratin type II cytoskeletal 6E Keratin-6C KRT6A KRT6B KRT6C KRT6D KRT6E type II......
Images
HA721456_1.jpg Fig1: Western blot analysis of Cytokeratin 5+6 on different lysates with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: NCI-H226 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 55 kDa

Exposure time: 1 minute;

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 (HA721456) 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.
HA721456_2.jpg Fig2: Western blot analysis of Cytokeratin 5+6 on different lysates with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/1,000 dilution.

Lane 1: Mouse brain tissue lysate (hot lysis)
Lane 2: Rat brain tissue lysate (hot lysis)
Lane 3: Rat brain tissue lysate (RIPA lysis)

Lysates/proteins at 40 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Exposure time: 1 minute 40 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 (HA721456) 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.
HA721456_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling Cytokeratin 5+6 with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721456_4.jpg Fig4: Immunocytochemistry analysis of A431 cells labeling Cytokeratin 5+6 with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 5+6 antibody (HA721456) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721456_5.jpg Fig5: Flow cytometric analysis of A431 cells labeling Cytokeratin 5+6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721456, 1ug/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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.