重组Anti-Collagen VI抗体[EPR17077] - C-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Collagen VI with ab199720 at 1/1600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on stromal cells of Human colon tissue is observed. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

AbReview77641****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed human palatine tonsil tissue. Stained with ab199720 at 1/500 dilution. Secondary antibody used was EnVision+ System- HRP Labelled Polymer anti rabbit. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% normal goat serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0

This image is courtesy of an anonymous Abreview

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Collagen VI with ab199720 at 1/1600 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Extracellular matrix staining on mouse cardiac muscle tissue is observed. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Blocking/dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the following literature : PMID : 16130093 and 21186846

All lanes:

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/10000 dilution

Lane 1:

Human fetal heart at 20 µg

Lane 2:

Human skeletal muscle at 20 µg

Lane 3:

WI-38 (Human lung) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 109 kDa

Observed band size: 147 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Lanes 1 - 3 : Merged signal (red and green). Green - ab199720 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab199720 was shown to recognize Collagen VI in wild-type Hek 293T cells as signal was lost at the expected MW in COL6A1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and COL6A1 knockout samples were subjected to SDS-PAGE. ab199720 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T whole cell lysate at 20 µg

Lane 2:

COL6A1 knockout HEK-293T whole cell lysate at 20 µg

Lane 3:

Human Skeletal Muscle whole cell lysate at 20 µg

Predicted band size: 109 kDa

false

• WB

Supplier Data

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (AB199720)

Blocking/dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the following literature : PMID : 16130093 and 21186846

All lanes:

Western blot - Anti-Collagen VI antibody [EPR17077] - C-terminal (ab199720) at 1/1000 dilution

Lane 1:

Mouse heart at 10 µg

Lane 2:

Mouse kidney at 10 µg

Lane 3:

Mouse spleen at 10 µg

Lane 4:

Rat heart at 10 µg

Lane 5:

Rat kidney at 10 µg

Lane 6:

Rat spleen at 10 µg

Lane 7:

NIH/3T3 (mouse embryo) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 109 kDa

Observed band size: 147 kDa

false

Exposure time: 10s