Explore
Validate
Learn710182
antibody from Invitrogen Antibodies
Targeting: CDKN1A
CAP20, CDKN1, CIP1, P21, p21CIP1, p21Cip1/Waf1, SDI1, WAF1
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 710182 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- p21 Recombinant Polyclonal Antibody (2HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 2HCLC
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing ofp21 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR865095_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of p21 was performed by loading 30 µg of MCF-7 wild type (Lane 1), MCF-7 Cas9 (Lane 2) and MCF-7 Cas9 cells transduced with p21 Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with p21 Recombinant Polyclonal Antibody (2HCLC) (Product # 710182, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL 1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific top21 (Fig (b)). Uncharacterized bands were observed t ~50 kDa in all the samples.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of p21 in whole cell extracts of MCF-7 using a p21 Recombinant Rabbit Polyclonal Antibody (Product # 710182) at a dilution of 1 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~21kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of p21 in HeLa cells using a p21 Recombinant Rabbit Polyclonal Antibody (Product # 710182) followed by detection using an Alexa Fluor 488-conjugated Goat anti-Rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of p21.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of p21 was done on 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with p21 Recombinant Rabbit Polyclonal Antibody (Product # 710182) at 1 µg\mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing nuclear localization of p21. Panel e shows no primary antibody. The images were captured at 20X magnification.
