Anti-Integrin beta 1 antibody [EP1041Y]

• IHC-P

AbReview9866****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Unpurified ab52971 staining human breast cancer metastasis tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with a commercial blocking reagent and incubation with the antibody (diluted 1/100) for 18 hours at 4°C. A HRP-conjugated goat anti-rabbit was used as the secondary antibody. This image shows a cancer metastasis at 40x with beta1 staining (in red) in both blood vessels and tumour cells. Blue is Hoechst for nuclei. The antibody detection was enhanced using a commercial Cy3 tyramide signal amplification kit.

This image is courtesy of an anonymous Abreview

• IHC-P

AbReview13423****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Formaldehyde-fixed, paraffin-embedded human lung tissue stained for Integrin beta 1 using ab52971 at 1/100 dilution in immunohistochemical analysis.

This image is courtesy of an Abreview submitted by Mark Southwood.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

ab52971 at 1/500 staining Integrin beta 1 antibody in human transitional cell carcinoma of bladder by immunohistochemistry (FFPE). Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling Integrin beta 1 with ab52971 at 1/500 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Lanes 1, 5 and 9 : Wild-type HAP1 cell lysate (20 μg)
Lanes 2, 6 and 10 : Integrin beta 1 knockout HAP1 cell lysate (20 μg)
Lanes 3, 7 and 11 : U87-MG cell lysate (20 μg)
Lanes 4, 8 and 12 : A431 cell lysate (20 μg)
Lanes 1, 2, 3 and 4 : Green signal from target – ab52971 observed at 140 kDa
Lanes 5, 6, 7 and 8 : Red signal from loading control – ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12 : Merged (red and green) signal

ab52971 was shown to specifically react with Integrin beta 1 in wild-type HAP1 cells. No band was observed when Integrin beta 1 knockout samples were examined. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab52971 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (ab52971)

Predicted band size: 88 kDa

false

• WB

Unknown

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

5% NFDM/TBST dilution buffer

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 1/10000 dilution

Lane 1:

U87-MG cell lysate at 20 µg

Lane 2:

HT-1080 cell lysate at 20 µg

Lane 3:

U937 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 88 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52971 overnight at 4°C. Antibody binding was detected using ab175781 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

Secondary antibody - anti-rabbit Alexa Fluor 790. ab175781.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 20 µg

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

HT 1080 (Human fibrosarcoma) Whole Cell Lysate at 20 µg

Lane 3:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-alexa-fluor-790-ab175781'>ab175781</a>) at 1/10000 dilution

Predicted band size: 88 kDa

Observed band size: 120 kDa,140 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Integrin beta 1 knockout HAP1 cell lysate (20 μg)
Lane 3 : U87-MG cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab52971 observed at 140 kDa. Red signal from loading control – ab8245 observed at 37 kDa.

This western blot image is a comparison between ab52971 and a competitor's top cited rabbit polyclonal antibody.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (ab52971)

Predicted band size: 88 kDa

false

• IHC

CiteAb

Immunohistochemistry - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Immunohistochemistry-immunofluorescence using Anti-Integrin beta 1 antibody [EP1041Y], ab52971. Publication image from Schuchman, E. H. et al., 2017, Cell Host Microbe, 28552668. Legend direct from paper.

Ectopic Expression of β1-Integrins Causes Acid Ceramidase Downregulation in Human Airway CF Cells(A–C) Ectopic surface expression of β1-integrin is found on the luminal pole of human CF bronchial epithelial cells (A), epithelial cells of nasal polyps (B), and freshly isolated nasal epithelial cells (C) but is absent from the luminal side of epithelial cells from healthy persons. Incubation with RGD peptides, anti-β1-integrin (Int) antibodies (HUTS-4), or acid ceramidase (Ac) normalized expression of β1-integrin in CF nasal epithelial cells (C). Sections from human CF or donor lungs or nasal polyps or freshly isolated, intact nasal epithelial cells were stained with Cy3-coupled anti-β1-integrin antibodies (Abcam) and analyzed by confocal microscopy. Shown are representative results from each of five CF or four healthy individuals (A and C) or each of four CF or healthy persons (B).Ac expression is downregulated in CF bronchial and nasal epithelial cells compared to healthy controls, (D) and (E). Normalization of ectopic β1-integrin expression resulted in re-expression of Ac (E). Fluorescence intensities in the luminal membrane were quantified using ImageJ and are given in arbitrary units (a.u.). Mean ± SD; n = 4 or 5; p values as given; t test for comparison of two values and ANOVA and post hoc Student’s t tests for all pairwise comparisons, applying Bonferroni correction for multiple comparisons. Overall p value for ANOVA is < 0.001 in all panels. The luminal membrane is indicated by arrows; E = epithelial cell layer. Please see also Figure S7.

• WB

CiteAb

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Western Blotting using Anti-Integrin beta 1 antibody [EP1041Y], ab52971. Publication image from Wu, M. et al., 2018, Nat Commun, 29335551. Legend direct from paper.

Exosomal miR-1247-3p activates fibroblasts via B4GALT3–β1-integrin–NF-κB signaling axis. a, b Immunoblotting assays of indicated proteins in MRC5 treated with CM or exosomes from different tumor cells. c, d Relative luciferase activity of NF-κB in MRC5 in the presence of CM or exosomes from different tumor cells. All data are shown as mean ± s.d. Student’s t-test was used to analyze the data. (*p < 0.05; **p < 0.01; ***p < 0.001). e Immunoblotting assays of indicated proteins in MRC5 with indicated treatments. f MiR-1247-3p effect on indicated proteins expression in MRC5 with or without suppression of β1-integrin expression. g Degradation assay of β1-integrin in MRC5 transfected with B4GALT3 or not

false

• WB

CiteAb

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Western Blotting using Anti-Integrin beta 1 antibody [EP1041Y], ab52971. Publication image from Wu, M. et al., 2018, Nat Commun, 29335551. Legend direct from paper.

Exosomal miR-1247-3p activates fibroblasts via B4GALT3–β1-integrin–NF-κB signaling axis. a, b Immunoblotting assays of indicated proteins in MRC5 treated with CM or exosomes from different tumor cells. c, d Relative luciferase activity of NF-κB in MRC5 in the presence of CM or exosomes from different tumor cells. All data are shown as mean ± s.d. Student’s t-test was used to analyze the data. (*p < 0.05; **p < 0.01; ***p < 0.001). e Immunoblotting assays of indicated proteins in MRC5 with indicated treatments. f MiR-1247-3p effect on indicated proteins expression in MRC5 with or without suppression of β1-integrin expression. g Degradation assay of β1-integrin in MRC5 transfected with B4GALT3 or not

false

• WB

CiteAb

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] (AB52971)

Western Blotting using Anti-Integrin beta 1 antibody [EP1041Y], ab52971. Publication image from Wu, M. et al., 2018, Nat Commun, 29335551. Legend direct from paper.

Exosomal miR-1247-3p activates fibroblasts via B4GALT3–β1-integrin–NF-κB signaling axis. a, b Immunoblotting assays of indicated proteins in MRC5 treated with CM or exosomes from different tumor cells. c, d Relative luciferase activity of NF-κB in MRC5 in the presence of CM or exosomes from different tumor cells. All data are shown as mean ± s.d. Student’s t-test was used to analyze the data. (*p < 0.05; **p < 0.01; ***p < 0.001). e Immunoblotting assays of indicated proteins in MRC5 with indicated treatments. f MiR-1247-3p effect on indicated proteins expression in MRC5 with or without suppression of β1-integrin expression. g Degradation assay of β1-integrin in MRC5 transfected with B4GALT3 or not

false