MA1-10808

antibody from Invitrogen Antibodies

Targeting: HBB beta-globin, CD113t-C

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-10808

Provider

Invitrogen Antibodies

Product name

Hemoglobin Monoclonal Antibody (HB11-231.2)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA1-10808 detects Hemoglobin from human samples. This antibody does not detect bovine, chicken, equine, ovine, or porcine Hemoglobin. MA1-10808 has been successfully used in ELISA and Western blot applications. The MA1-10808 immunogen is purified human hemoglobin.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

HB11-231.2

Vial size

500 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Hemoglobin Monoclonal Antibody (HB11-231.2) (Product # MA1-10808) and a 15 kDa band corresponding to Hemoglobin was observed only in K-562 which is reported to be positive and not in any other cell lines. Whole cell extracts (30 µg lysate) of K-562 (Lane 1), HL-60 (Lane 2), Raji (Lane 3), U-2 OS (Lane 4) and A549 (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 (2 ug/ml) 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Hemoglobin was performed using K-562 cells. 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 Hemoglobin Monoclonal Antibody (HB11-231.2) (Product # MA1-10808) at 1:100 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: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents HL-60 cells having no expression of Hemoglobin. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.