All lanes : Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248) at 1 µg/ml

Lane 1 : Hek293
Lane 2 : HL60
Lane 3 : HeLa
Lane 4 : A549
Lane 5 : Hu spleen
Lane 6 : Ms spleen
Lane 7 : Rt spleen

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : IRDye® 800CW Goat anti Mouse

Predicted band size: 34.6 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?

Hek293 & HL60 presumed negative or very low expression.

Loading control GAPDH at 38kDa

This image was generated using the ascites version of the product.

IHC image of Hem Oxygensae 1 staining in a section of formalin fixed, paraffin embedded normal human spleen tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab13248, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

This image was generated using the ascites version of the product.

 

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

The following proteins and lysates were electrophoresed; Lane 1 - Heme-Oxygenase-1 (Hsp32) Protein (50ng), lane 2 - Heme-Oxygenase-2 protein NSP-550 (100ng), lane 3 - MDBK Cell Lysate (20ug), lane 4 - Mouse liver microsome (20ug) and lane 5 - Dog liver microsome (20ug). ab13248 was applied at a concentration of 4ugml-1.

This image was generated using the ascites version of the product.

Standard Curve for Heme Oxygenase 1 (Analyte: Heme Oxygenase 1 protein (Tagged) (ab85243)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [HO-1-1] to Heme Oxygenase 1 (ab13248) at 5µg/ml and Detector Antibody Rabbit polyclonal to Heme Oxygenase 1 (ab13243) at 0.5µg/ml.

This image was generated using the ascites version of the product.

Lane 1 : MW marker
Lanes 2-7 : Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248)

Lane 2 : Recombinant Rat Heme Oxygenase 1
Lane 3 : Recombinant Human Heme Oxygenase 1
Lane 4 : Recombinant Human Heme Oxygenase 2
Lane 5 : MDBK cell lysate
Lane 6 : Dog liver microsome
Lane 7 : Mouse liver microsome

Predicted band size: 34.6 kDa

This image was generated using the ascites version of the product.

Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248) at 1/250 dilution + Human microsome lysate

Predicted band size: 34.6 kDa

This image was generated using the ascites version of the product.

ab13248 at 10µg/ml staining Heme Oxygenase 1 in human lung cancer A2 cells by flow cytometery. The left image repersents staining with isotype control antibody and the right image show staining with ab13248.

This image was generated using the ascites version of the product.