Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA5-20289 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SLUG Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control is human kidney tissue lysate.
- Concentration
- 1 mg/mL
Submitted references Parathyroid Hormone-Related Protein Inhibition Blocks Triple-Negative Breast Cancer Expansion in Bone Through Epithelial to Mesenchymal Transition Reversal.
Li J, Camirand A, Zakikhani M, Sellin K, Guo Y, Luan X, Mihalcioiu C, Kremer R
JBMR plus 2022 Jun;6(6):e10587
JBMR plus 2022 Jun;6(6):e10587
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of human kidney cell lysate using a Slug polyclonal antibody (Product # PA5-20289) at in (A) 1 and (B) 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of Slug in human kidney cell lysate with SLUG Polyclonal Antibody (Product # PA5-20289) at in (A) 1 and (B) 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SLUG Polyclonal Antibody (Product # PA5-20289) and a 29kDa band corresponding to Slug was observed across all the cell lines tested and increased upon TGF-Beta treatment in MCF7. Whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), MDA-MB-231 (Lane 2), MCF-7 (Lane 3), MCF-7 treated with TGF-Beta (10ng/ml for 4 days (Lane 4), MCF 10A (Lane 5), U-937 (Lane 6) and Daudi were electrophoresed using NuPAGE™ 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 the primary antibody (1µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SLUG was performed using 70% confluent log phase MCF-7 cells treated with 10 ng/mL TGF-Beta for 4 days. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled Anti-SLUG Polyclonal Antibody (Product # PA5-20289) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034) for 45 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents untreated MCF-7 cells having low expression of SLUG. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
