p21 Recombinant Rabbit Monoclonal Antibody [PSH22-37]
cat.: HA724295
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, IP
Clonality: Monoclonal
Clone number: PSH22-37
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: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 18 kDa
Isotype: IgG
Positive control: HCT 116 cell lysate, A549 cell lysate, MCF7 cell lysate, MCF7 treated with 10μM Nutlin 3a for 24 hours cell lysate, MCF7, human colon cancer tissue, human ovarian cancer tissue, human breast cancer tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IP
1:2,000
1:5,000
1:200
1-2μg/sample
Uniprot #: SwissProt: P38936 Human
Alternative names: CAP20 CDK-interacting protein 1 CDKI CDKN1 Cdkn1a CDN1A_HUMAN CIP1 Cyclin Dependent Kinase Inhibitor 1A Cyclin-dependent kinase inhibitor 1 Cyclin-dependent kinase inhibitor 1A (P21) Cyclin-dependent kinase inhibitor 1A (p21, Cip1) DNA Synthesis Inhibitor MDA-6 MDA6 Melanoma differentiation-associated protein 6 Melanoma differentiation-associated protein p21 P21 protein p21CIP1 p21Cip1/Waf1 p21WAF PIC1 SDI1 SLC12A9 WAF1 Wild type p53 activated fragment 1 (WAF1) Wild type p53 activated fragment 1 Wildtype p53-activated fragment 1
Images
HA724295_1.jpg Fig1: Western blot analysis of p21 on different lysates with Rabbit anti-p21 antibody (HA724295) at 1/2,000 dilution.

Lane 1: HCT 116 (Human colon cancer cells)
Lane 2: NCI-H1299 (Human non-small cell Lung carcinoma cells) (Negative)
Lane 3: A549 (Human lung adenocarcinoma cells)
Lane 4: MCF7 (Human breast cancer cells)
Lane 5: MCF7 (Human breast cancer cells) treated with Nutlin 3a (10uM, 24h)

Lysates/proteins at 10 µg/Lane.
Exposure time: 1 minutes; ECL: K1801

H1299 is a negative control (PMID: 23268708).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724295, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 21 kDa
Observed band size: 21 kDa
HA724295_2.jpg Fig2: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: MCF7 (Human cervix adenocarcinoma epithelial cell) treated or untreated with Nutlin 3a (10 μM, 24h)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724295, 1/5,000, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
HA724295_3.jpg Fig3: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Colon cancer
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× PBST
Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724295, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724295_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Ovarian Carcinoma
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× PBST
Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724295, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724295_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Invasive Ductal Carcinoma of the Breast
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× PBST
Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724295, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724295_6.jpg Fig6: Application: Immunoprecipitation (IP)

p21 was immunoprecipitated in 0.2 mg HCT 116 (Human colon cancer cell) cell lysate with HA724295 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724295 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HCT 116 cell lysate (input)
Lane 2: Rabbit IgG instead of HA724295 in HCT 116 cell lysate
Lane 3: HA724295 IP in HCT 116 cell lysate

Exposure time: 1 minute
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724295, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 21 kDa
Observed band size: 21 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.