- PA1-064 - Invitrogen Antibodies CAV1 antibody

Submitted references Compression enhances invasive phenotype and matrix degradation of breast Cancer cells via Piezo1 activation.

Luo M, Cai G, Ho KKY, Wen K, Tong Z, Deng L, Liu AP
BMC molecular and cell biology 2022 Jan 3;23(1):1

Involvement of the TGF-β1 pathway in caveolin-1-associated regulation of head and neck tumor cell metastasis.

Sun J, Lu Y, Yu C, Xu T, Nie G, Miao B, Zhang X
Oncology letters 2020 Feb;19(2):1298-1304

Occludin, caveolin-1, and Alix form a multi-protein complex and regulate HIV-1 infection of brain pericytes.

Torices S, Roberts SA, Park M, Malhotra A, Toborek M
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2020 Dec;34(12):16319-16332

Energetic costs regulated by cell mechanics and confinement are predictive of migration path during decision-making.

Zanotelli MR, Rahman-Zaman A, VanderBurgh JA, Taufalele PV, Jain A, Erickson D, Bordeleau F, Reinhart-King CA
Nature communications 2019 Sep 13;10(1):4185

Aroclor1254 disrupts the blood-testis barrier by promoting endocytosis and degradation of junction proteins via p38 MAPK pathway.

Jia X, Xu Y, Wu W, Fan Y, Wang G, Zhang T, Su W
Cell death & disease 2017 May 25;8(5):e2823

Colocalization of androgen binding protein, oxytocin receptor, caveolin 1 and proliferation marker p21 in benign prostate hyperplasia.

Sendemir E, Herbert Z, Sivukhina E, Zermann DH, Arnold R, Jirikowski GF
Anatomia, histologia, embryologia 2008 Oct;37(5):325-31

Expression of sex hormone-binding globulin, oxytocin receptor, caveolin-1 and p21 in leiomyoma.

Sendemir A, Sendemir E, Kosmehl H, Jirikowski GF
Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology 2008 Feb;24(2):105-12

Changing caveolin-1 and oxytocin receptor distribution in the ageing human prostate.

Herbert Z, Bötticher G, Aschoff A, Sendemir E, Zermann DH, Arnold R, Mall G, Jirikowski GF
Anatomia, histologia, embryologia 2007 Oct;36(5):361-5

Signaling proteins in raft-like microdomains are essential for Ca2+ wave propagation in glial cells.

Weerth SH, Holtzclaw LA, Russell JT
Cell calcium 2007 Feb;41(2):155-67

Isolation of lipid droplets from cells by density gradient centrifugation.

Brasaemle DL, Wolins NE
Current protocols in cell biology 2006 Jan;Chapter 3:Unit 3.15

A high-fat, refined-carbohydrate diet induces endothelial dysfunction and oxidant/antioxidant imbalance and depresses NOS protein expression.

Roberts CK, Barnard RJ, Sindhu RK, Jurczak M, Ehdaie A, Vaziri ND
Journal of applied physiology (Bethesda, Md. : 1985) 2005 Jan;98(1):203-10

Effects of chronic renal failure on caveolin-1, guanylate cyclase and AKT protein expression.

Sindhu RK, Ehdaie A, Vaziri ND, Roberts CK
Biochimica et biophysica acta 2004 Nov 5;1690(3):231-7

Decreased liver fatty acid binding capacity and altered liver lipid distribution in mice lacking the liver fatty acid-binding protein gene.

Martin GG, Danneberg H, Kumar LS, Atshaves BP, Erol E, Bader M, Schroeder F, Binas B
The Journal of biological chemistry 2003 Jun 13;278(24):21429-38

Cell-specific targeting of caveolin-1 to caveolae, secretory vesicles, cytoplasm or mitochondria.

Li WP, Liu P, Pilcher BK, Anderson RG
Journal of cell science 2001 Apr;114(Pt 7):1397-408

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of PANC-1 (Lane 1), U-87 MG (Lane 2), A549 (Lane 3), HeLa (lane4), PC-3 (lane 5), U2OS (lane 6), Mouse Heart (lane 7), Rat Heart (lane 8), C2C12 (lane 9), Mouse Lung (lane 10) and A-431 (lane 11). The blots were probed with Anti-Caveolin-1 Rabbit Polyclonal Antibody (Product # PA1-064, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 17 kDa band corresponding to Caveolin-1 was observed across cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of PANC-1 (Lane 1), U-87 MG (Lane 2), A549 (Lane 3), HeLa (lane4), PC-3 (lane 5), U2OS (lane 6), Mouse Heart (lane 7), Rat Heart (lane 8), C2C12 (lane 9), Mouse Lung (lane 10) and A-431 (lane 11). The blots were probed with Anti-Caveolin-1 Rabbit Polyclonal Antibody (Product # PA1-064, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 17 kDa band corresponding to Caveolin-1 was observed across cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Caveolin 1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR639775_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Caveolin 1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Wild Type treated with Palcitaxel (25 nM for 24 hrs) (Lane 2), HeLa Cas9 (Lane 3), HeLa Cas9 treated with Palcitaxel (25 nM for 24 hrs) (Lane 4), HeLa Caveolin 1 KO (Lane 5) and HeLa Caveolin 1 KO treated with Palcitaxel (25 nM for 24 hrs) (Lane 6) 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 Anti-Caveolin 1 Polyclonal Antibody (Product # PA1-064, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:2,500 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Caveolin 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Caveolin 1 was done on 70% confluent log phase A-375 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Caveolin 1 Rabbit Polyclonal Antibody (Product # PA1-064) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Caveolin 1 was done on U-87 MG cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Caveolin 1 Rabbit Polyclonal Antibody (PA1064, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Caveolin 1 was immunoprecipitated using 5 µg of the Caveolin 1 Rabbit Polyclonal Antibody (Product # PA1-064) from lysate of Mouse Heart (Lane 3) using the Dynabeads® Protein A Immunoprecipitation Kit (Product # 10006D). Normal Rabbit IgG was used as a Isotype control (Lane 2). 10 % input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using Caveolin 1 Rabbit Polyclonal Antibody (Product # PA1-064) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Caveolin 1 was immunoprecipitated using 5 æg of the Caveolin 1 Rabbit Polyclonal Antibody (Product # PA1-064) from lysate of Mouse Heart (Lane 3) using the Dynabeads® Protein A Immunoprecipitation Kit (Product # 10006D). Normal Rabbit IgG was used as a Isotype control (Lane 2). 10 % input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using Caveolin 1 Rabbit Polyclonal Antibody (Product # PA1-064) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 æg/mL, 1:2500 dilution). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 1 Prostate stroma of a BPH patient. Staining with anti-ABP (a), anti-Caveolin-1 (b), anti-Oxytocin receptor (c) or anti-p21 (d) antibodies, developed by DAB. Note the ABP immunoreactivity in a prostate stone (e). Serial semithin sections (1 mu m thick) of the prostate tissue. Scale bar indicates 10 mu m (a-d) and 100 mu m for (e).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 2 Serial semithin sections of a prostate control sample. Immunohistochemical staining with anti-ABP (a), anti-Caveolin-1 (b), anti-Oxytocin receptor (c) or anti-p21 (d) antibodies, developed by DAB. Scale bar indicates 10 mu m (a-d).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 HIV-1 infection alters cav-1, ocln, and Alix expression in brain pericytes. Pericytes were either mock-infected or infected with 60 ng/mL HIV-1 p24 for 48 h (A) or 72 h (B). The expression of cav-1, ocln, and Alix was evaluated by immunoblotting. GAPDH was used as a loading control. C, Representative immunostaining images of cav-1 (green), ocln (red), and Alix (purple) in mock-infected or HIV-1-infected for 48 or 72 h. Alix expression intensity was increased by the same factors in all groups to allow for better visualization. Graphs show the mean +- SD from three independent experiments. **** P < .0001, ** P = .003, * P < .0449, n = 4-9 per group; scale bars, 20 um

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Cav-1, ocln, and Alix form a stable complex in mock-infected and HIV-1-infected pericytes. A, Diagram illustrating Alix and its binding partners identified in this study in a structural ribbon representation. The ESCRT-associated protein Alix binds tyrosine motifs via its C-terminal Proline Rich Domain (PRD). Caveolae are made up of oligomers of cav-1 and cavin proteins, and the figure shows a trimeric coiled coil for Cavin4a HR1 domain. For ocln, the region shown is the cytoplasmic C-terminal region that is known to bind scaffolding proteins. Figure was prepared using PyMol with the listed PDB accession numbers. B-E, Pericytes were either mock-infected (B) or infected with HIV-1 (C) for 48 h as in Figure 1 . A total of 600 ug of cell lysate protein was incubated with cav-1 antibody for 24 h, followed by incubation with protein A/G PLUS-Agarose beads. Both immunoprecipitates and supernatants were analyzed by immunoblotting for the presence of cav-1, ocln, and Alix. D, Immunostaining of cav-1 (green), ocln (red), and Alix (purple) in mock-infected or HIV-1-infected for 48 h. E, Cellular localization of the cav-1, ocln, and Alix complex in mock-infected or HIV-1-infected for 48 h. Cell membranes are visualized by blue fluorescent dye DiB (left panel). Middle panel, the cav-1, ocln, and Alix complexes merged with a bright field image of a single pericyte. The images were analyzed as in Figure 1 ; n = 4 per group; scale bar 5, 10, or 20 um

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 3 HIV-1 infection attenuates cav-1-mediated regulation of ocln expression. Pericytes were transfected with 1 ug cav-1 siRNA per 10 6 cells and either mock-infected or HIV-1-infected with 60 ng/mL HIV-1 p24 for 48 h. The expression of cav-1, ocln, and Alix was evaluated by immunoblotting (A), quantified, and compared among groups. GAPDH was used as a loading control. Cav-1 silencing (B) resulted in upregulation of occludin levels in mock-infected pericytes and this effect was attenuated in HIV-1-infected pericytes (C). Cav-1 silencing did not affect Alix levels (D). Graphs indicate the mean +- SD from three independent experiments. **** P < .0001, *** P = .0002, ** P = .003, n = 4-9 per group

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Cell stiffness alters decision-making and cell mechanics during confined migration. a AFM measurements of cell stiffness following pharmacological treatments and siCav1 knockdown (n= 164, 95, 167, 91, 75, 85, 92, and 97 cells for Ctrl, Rho+, CL-A, Y27, ML7, MßCD, siCtrl, and siCav1) and western blot for Caveolin-1 following treatment with 25 nM Caveolin-1 siRNA. b, c Migration decision-making (b) and passing time into each branch (c) following treatments to alter cell stiffness (n= 30, 34, 30, 31, 30, 28, 33, and 39 cells for Ctrl, Rho+, CL-A, Y27, ML7, MßCD, siCtrl, and siCav1). d-f Model predictions for cell deformation (d), track displacement (e), and the inverse relationship between cell elongation and track displacement (f) in 7 µm tracks. g Transmitted light images showing cell morphology and confocal reflectance images of microtrack structure as well as cell-induced track displacement in 7 µm microtracks (yellow lines show microtrack walls; red arrowheads show areas of matrix displacement around the cell body). h-j Experimental averages of cell elongation (h), track displacement (i), and the relationship between cell remodeling and matrix displacement (j) in 7 µm microtracks as a function of cell stiffness (n= 118, 107, 100, 135, 109, 123, 53, and 58 cells for Ctrl, Rho+, CL-A, Y27, ML7, MßCD, siCtrl, and siCav1). Data shown as median interquartile range (box), 5th-95th percentiles (whiskers), and mean (+) (a, c), or mean s.e.m. (b, h-j); dashed lines show one-

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. Caveolin-1 knockdown in Tu686 cells by shRNA. (A) Western blot analysis depicting the reduced levels of caveolin-1 following knockdown by shRNA. The levels of caveolin-1 in the interfered group1 were significantly inhibited. Data were analyzed using the Student's t-test. *P

PA1-064

Antibody data