- MA5-29035 - Invitrogen Antibodies APOH antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-Apolipoprotein H Monoclonal Antibody (4A2F8H12) (Product # MA5-29035) and a 50 kDa band corresponding to Beta-2-glycoprotein 1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with PTI (1X for 4 hours) (Lane 2), PC-3 (Lane 3), PC-3 treated with PTI (1X for 4 hours) (Lane 4), BeWo (Lane 5), BeWo treated with PTI (1X for 4 hours) (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence staining of Human Apolipoprotein H in HepG2 or Hela cells. Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with Apolipoprotein H Monoclonal Antibody (4A2F8H12) (Product # MA5-29035, 1:60). Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (left panel, captured by laser confocal scanning microscope; right panel, captured by fluorescence microscope), counterstained with DAPI (blue). Positive staining was localized to cytoplasm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Beta-2-glycoprotein 1 was performed using 70% confluent log phase Hep G2, Hep G2 treated with PTI (1X for 4hrs) and PC-3 cells treated with PTI (1X for 4hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Apolipoprotein H Monoclonal Antibody (4A2F8H12) (Product # MA5-29035) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), for 45 minutes at room temperature (Panel a,e,i : Green). Nuclei (Panel b,f,j :Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c,g,k : Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel h represents the merged image showing cytoplasmic localization upon PTI treatment. Panel d and i represents merged images for control cells. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of Human Apolipoprotein H expression on MDA-MB-231 cells. The cells were treated according to manufacturer’s manual, stained with Apolipoprotein H Monoclonal Antibody (4A2F8H12) (Product # MA5-29035), then a FITC-conjugated Secondary antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.

MA5-29035