Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA1-37934 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Involucrin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is skin tissue.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 1 mL
- Concentration
- 0.267 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Essential Functions of Glycans in Human Epithelia Dissected by a CRISPR-Cas9-Engineered Human Organotypic Skin Model.
A novel gain-of-function mutation of TGF-β receptor II promotes cancer progression via delayed receptor internalization in oral squamous cell carcinoma.
Dabelsteen S, Pallesen EMH, Marinova IN, Nielsen MI, Adamopoulou M, Rømer TB, Levann A, Andersen MM, Ye Z, Thein D, Bennett EP, Büll C, Moons SJ, Boltje T, Clausen H, Vakhrushev SY, Bagdonaite I, Wandall HH
Developmental cell 2020 Sep 14;54(5):669-684.e7
Developmental cell 2020 Sep 14;54(5):669-684.e7
A novel gain-of-function mutation of TGF-β receptor II promotes cancer progression via delayed receptor internalization in oral squamous cell carcinoma.
Park I, Son HK, Che ZM, Kim J
Cancer letters 2012 Feb 28;315(2):161-9
Cancer letters 2012 Feb 28;315(2):161-9
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Involucrin Polyclonal Antibody (Product # PA5-32454) and ~68kDa, ~75kDa, ~100kDa, ~124kDa bands corresponding to Involucrin was observed across skin derived cell lines tested (HaCaT and A-431) and not in others. Whole Cell Extract (30 µg lysate) of HaCaT (Lane 1), A-431 (Lane 2), HeLa (Lane 3), MDA-MB-231 (Lane 4), Raji (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). IVL is reported to show this pattern of expression at molecular weights >68kDa and all of these forms are not detected in the low and/or negative cell lines tested [doi:10.1242/jcs.056093; 10.1093/toxsci/kfm153]
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Involucrin was performed using 70% confluent log phase HaCaT and MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 45 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Involucrin Polyclonal Antibody (Product # PA5-32454) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization of IVL in HaCaT. Panel e represents MDA-MB-231 cells showing no staining for IVL. Panel f represents control HaCaT cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Involucrin using anti-Involucrin Polyclonal Antibody (Product # PA5-32454) in Skin Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.
