Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Human breast tissue labelling Fibrillin 1 with ab315384 at 1 : 100 dilution (B), Cytokeratin 19 with ab52625 at 1 : 6000 dilution (C) and Hormone sensitive lipase/HSL with ab322344 at 1 : 2000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Fibrillin 1 (green; Opal™570), anti-Cytokeratin 19 (magenta; Opal™690) and anti-Hormone sensitive lipase/HSL (gray; Opal™570) on human breast.

Panel B : anti-Fibrillin 1 staining stromal connective tissue in human breast.

Panel C : ant-Cytokeratin 19 staining epithelium in human breast.

Panel D : ant-Hormone sensitive lipase/HSL staining adipocytes in human breast.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315384, ab52625 and ab322344 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal connective tissue of human colon carcinoma. The section was incubated with ab315384 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with citrate buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-937 (human histiocytic lymphoma monocyte, Left) / A-172 (human brain glioblastoma cell, Right) cells labelling Fibrillin 1 with ab315384 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : U-937.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal connective tissue of human testis. The section was incubated with ab315384 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with citrate buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal connective tissue of human hepatocellular carcinoma. The section was incubated with ab315384 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with citrate buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal connective tissue of human skin (PMID : 16120151, PMID : 20195245). The section was incubated with ab315384 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with citrate buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A-172 (human brain glioblastoma cell) cells labelling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in A-172 cell line. Negative control : U-937. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ARPE-19 (human eye epithelial cell) cells labelling Fibrillin 1 with ab315384 at 1/100 (5.08 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in ARPE-19 cell line. Negative control : T-47D. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized T-47D (human ductal breast epithelial tumor epithelial cell, Left) / ARPE-19 (human eye epithelial cell, Right) cells labelling Fibrillin 1 with ab315384 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : T-47D.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab315384 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Human Fibrillin-1,Human Fibrillin-2,Human Fibrillin-3.

Antigen concentration : 1000 ng/ml

• Dot

Supplier Data

Dot Blot - Anti-Fibrillin 1 antibody [EPR27362-78] - BSA and Azide free (AB315385)

This data was developed using ab315384, the same antibody clone in a different buffer formulation.

Dot blot analysis of Fibrillin 1 using ab315384 at 1 : 1000 (0.508 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 1000 (0.508 ug/ml) dilution.

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody has weak cross reaction with human Fibrillin 3.

The recombinant proteins of Fibrillin 1, Fibrillin 2 and Fibrillin 3 were made in house.