Anti-MMP1 antibody [RM1206] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MMP1 antibody [RM1206] - BSA and Azide free (AB318208)

This data was developed using ab318207, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling MMP1 with ab318207 at 1/500 (0.986 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 and membranous staining in HUVEC cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Low expression : A549.
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 (Magenta). 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.

• IP

Supplier Data

Immunoprecipitation - Anti-MMP1 antibody [RM1206] - BSA and Azide free (AB318208)

This data was developed using ab318207, the same antibody clone in a different buffer formulation.

MMP1 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab318207 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318207 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 2 : ab318207 IP in HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318207 in HUVEC whole cell lysate

All lanes:

Immunoprecipitation - Anti-MMP1 antibody [RM1206] (<a href='/en-us/products/primary-antibodies/mmp1-antibody-rm1206-ab318207'>ab318207</a>) at 1/30 dilution

All lanes:

HUVEC (human umbilical vein endothelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-MMP1 antibody [RM1206] - BSA and Azide free (AB318208)

This data was developed using ab318207, the same antibody clone in a different buffer formulation.

Low expression : A549, HT-29, IMR-32.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 25319362).

Anti-MMP1 antibody (ab318207, Left) staining at 1/1000 dilution showed similar expression profile/ molecular weight with Anti-MMP1 antibody (ab137332, Right) at 1/1000 dilution.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MMP1 antibody [RM1206] (<a href='/en-us/products/primary-antibodies/mmp1-antibody-rm1206-ab318207'>ab318207</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

HeLa treated with 80 nM TPA for 24 hours, then replaced with 10 ug /ml poly(dA:dT) for 2 hours, whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 5:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 6:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 54 kDa,36 kDa

false

Exposure time: 180s