AM00301AF-N CDKN2C / p18INK4c antibody

See related secondary antibodies

Search for all "CDKN2C / p18INK4c"

0.1 mg / €355.00
Please visit the country specific website of OriGene Technologies or contact your local Distributor to buy this product.

Quick Overview

Mouse anti Human CDKN2C / p18INK4c DCS-118

AM00301AF-N

More Views

  • AM00301AF-N

Product Description for CDKN2C / p18INK4c

Mouse anti Human CDKN2C / p18INK4c DCS-118.
Presentation: Azide Free
Product is tested for Immunoprecipitation, Paraffin Sections, Western blot / Immunoblot.

Properties for CDKN2C / p18INK4c

Product Category Primary Antibodies
Target Category
Quantity 0.1 mg
Synonyms CDKN6, Cyclin-dependent kinase 4 inhibitor C, Cyclin-dependent kinase 6 inhibitor, p18-INK4c, p18-INK6, p18INK6
Presentation Azide Free
Reactivity Hu
Applications IP, P, WB
Clonality Monoclonal
Clone DCS-118
Host Mouse
Isotype IgG2a
Shipping to Not USA/Canada
PDF datasheet View Datasheet
Manufacturer OriGene Technologies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Swiss Prot Num:
P42773
Immunogen:
Bacterially produced His-tagged p18 proteins.
GeneID:
1031
Add. information This product was originally produced by MBL International.
Application Western Blot: 1 µg/mL
Positive Control: Saos-2 Cells.
Immunoprecipitation: 3 µg/200-300 µL of cell extract.
Positive Control: Saos-2 Cells.
Immunohistochemistry: 1-5 μg/mL
Heat treatment is necessary for Paraffin Embedded Sections.
Microwave oven: 2 times for 10 minutes each in citrate buffer (pH 6.5).
Positive Control: Tonsil Tissue.
Detailed procedure is provided in Protocols.
Background The INK4 family of proteins consists of four members that block progression from the G1-to-S phase of the cell cycle by inhibiting the activity of Cdk4 and Cdk6. The p18INK4c cyclin-dependent kinase inhibitor is an important regulator of cellular differentiation and cell cycle progression, and it also acts as a potent tumor suppressor. p18INK4c is regulated by the transcription factors E2F1 and SP1 in response to environmental and intracellular signals such as cytokines, oncogenic overload, or cellular senescence.
Concentration 1.0 mg/ml
Protocols SDS-PAGE & Western Blotting
1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds).
2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the Lysis buffer to make 8 mg/mL solution.
3) Mix the sample with equal volume of Laemmli’s sample buffer.
4) Boil the samples for 2 minutes and centrifuge. Load 10 μL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis.
5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hour in a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for the transfer procedure.
6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4°C.
7) Incubate the membrane with the anti-p18INK4c (DCS-118) monoclonal antibody (1 μg/mL) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature.
8) Wash the membrane with PBS (5 minutes x 6 times).
9) Incubate the membrane with the 1:10000 HRP-conjugated anti-mouse IgG diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature.
10) Wash the membrane with PBS (5 minutes x 6 times).
11) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap.
12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary.
Positive Control for Western blotting: Saos-2 cells.

Immunoprecipitation
1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds).
2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant to another tube.
3) Add 3 μg of the anti-p18INK4c (DCS-118) monoclonal antibody into 250 μL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4°C. Add 20 μL of 50% Protein A-agarose beads resuspended in the Lysis buffer. Mix well and incubate with gentle agitation for 60 minutes at 4°C.
4) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds).
5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis.
(See SDS-PAGE & Western blotting.)
Positive Control for immunoprecipitation: Saos-2 cells.


Immunohistochemical Staining for Paraffin-Embedded Sections: SAB method
1) Deparaffinize the sections with Xylene 3 times for 3-5 minutes each.
2) Wash the slides with Ethanol 3 times for 3-5 minutes each.
3) Wash the slides with PBS 3 times for 3-5 minutes each.
4) Heat treatment
Heat treatment by microwave oven: Place the slides put on staining basket in 500 mL beaker with 500 mL citrate buffer (pH 6.5). Cover the beaker with plastic wrap, then process the slides 2 times for 10 minutes each at 500 W with microwave oven. Let the slides cool down in the beaker at room temperature for about 40 minutes.
5) Remove the slides from the citrate buffer and cover each section with 3% H2O2 for 10 minutes at room temperature to block endogenous peroxidase activity. Wash 3 times in PBS for 5 minutes each.
6) Remove the slides from PBS, wipe gently around each section and cover tissues with Protein Blocking Agent for 5 minutes to block non-specific antibody staining. Do not wash.
7) Tip off the blocking buffer, wipe gently around each section and cover tissues with the anti-p18INK4c (DCS-118) monoclonal antibody diluted with PBS containing 1% BSA (1-5 µg/mL).
8) Incubate the sections for 1 hour at room temperature.
9) Wash the slides 3 times in PBS for 5 minutes each.
10) Wipe gently around each section and cover tissues with Polyvalent Biotinylated Antibody. Incubate for 10 minutes at room temperature. Wash as in step 9.
11) Wipe gently around each section and cover tissues with Streptavidin-Peroxidase. Incubate for 10 minutes at room temperature. Wash as in step 9.
12) Visualize by reacting for 10-20 minutes with substrate solution containing 7.5 mg DAB, 40 µL of 30% H2O2 in 150 mL PBS. *DAB is a suspected carcinogen and must be handled with care. Always wear gloves.
13) Wash the slides in water for 5 minutes.
14) Counter stain in hematoxylin for 1 minute, wash the slides 3 times in water for 5 minutes each, and then immerse the slides in PBS for 5 minutes. Dehydrate by immersing in Ethanol 3 times for 3 minutes each, followed by immersing in Xylene 3 times for 3 minutes each.
15) Now ready for mounting.
Positive Control for Immunohistochemistry: Tonsil Tissue.
General Readings 1. Thullberg, M., et al. Hybridoma 19, 63-72 (2000)
Clone DCS-118 is used in this reference.
Storage Store the antibody undiluted at -20°C.
Shelf life: one year from despatch.
Format
Purification:
Protein-A Sepharose Chromatography.
Buffer System:
PBS, pH 7.2 containing 50% Glycerol without preservatives.
State:
Liquid purified IgG fraction.
Azide Free
Species Reactivity
Species reactivity (tested):
Human.
Specificity
Specificity:
This antibody reacts with Human p18INK4c.
Gene ID 1031
FocusOn and Reviews
FocusOns:

Accessory Products

Proteins and/or Positive Controls

Proteins for CDKN2C / p18INK4c (7 products)

Catalog No. Species Pres. Purity   Source  

CDKN2C / p18INK4c (transcript variant 1)

CDKN2C / p18INK4c Human > 80 %
Preparation: Recombint protein was captured through anti-DDK affinity column followed by conventiol chromatography steps.
Purity Detail: > 80% as determined by SDS-PAGE and Coomassie blue staining.
HEK293 cells
20 µg / €748.00
  OriGene Technologies, Inc.

CDKN2C / p18INK4c (transcript variant 2)

CDKN2C / p18INK4c Human > 80 %
Preparation: Recombint protein was captured through anti-DDK affinity column followed by conventiol chromatography steps.
Purity Detail: > 80% as determined by SDS-PAGE and Coomassie blue staining.
HEK293 cells
20 µg / €748.00
  OriGene Technologies, Inc.

CDKN2C / p18INK4c (transcript variant 1)

CDKN2C / p18INK4c Human > 95 %
Preparation: .
Purity Detail: >95% as determined by SDS-PAGE and Coomassie blue staining.
E. coli
10 µg / €299.00
  OriGene Technologies, Inc.

CDKN2C / p18INK4c (1-168, His-tag)

CDKN2C / p18INK4c Human Purified > 95 % by SDS - PAGE E. coli
  OriGene Technologies GmbH

CDKN2C / p18INK4c (1-168, His-tag)

CDKN2C / p18INK4c Human Purified > 95 % by SDS - PAGE E. coli
  OriGene Technologies GmbH

CDKN2C / p18INK4c

CDKN2C / p18INK4c Human Purified 12.5% SDS-PAGE Stained with Coomassie Blue.
  Abnova Taiwan Corp.

CDKN2C / p18INK4c

CDKN2C / p18INK4c Human Purified 12.5% SDS-PAGE Stained with Coomassie Blue.
  Abnova Taiwan Corp.

Positive controls for CDKN2C / p18INK4c (2 products)

Catalog No. Species Pres. Purity   Source  

CDKN2C 293T Cell Transient Overexpression Lysate(Denatured)

CDKN2C 293T Cell Transient Overexpression Lysate(Denatured)
  Abnova Taiwan Corp.

p18 INK4c Lysate

Western Blot: p18 INK4c Lysate [NBL1-09055] - Western Blot experiments. 
Left-Control; Right -Over-expression Lysate for CDKN2C
  Novus Biologicals Inc.
  • LinkedIn