PA5-52765

antibody from Invitrogen Antibodies

Targeting: MFF C2orf33, GL004

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-52765

Provider

Invitrogen Antibodies

Product name

C2orf33 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: RFQAPISAPE YTVTPSPQQA RVCPPHMLPE DGANLSSARG ILSLIQSSTR RAYQQILDVL DENRRPVLRG GSAAATSNPH HDNVRYGISN IDTTIEGTSD DLTVVDAASL RRQIIKLNRR LQLLEEENKE RAKR

Concentration

0.1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

KD of C2orf33 was achieved by transfecting HeLa cells with C2orf33 specific siRNAs (Silencer® select Product # s25664, s25665). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the C2orf33 KD cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with C2orf33 Polyclonal Antibody (Product # PA5-52765, 1:1000 dilution) and Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to C2orf33.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-C2orf33 Rabbit Polyclonal Antibody (Product # PA5-52765) and 28kDa, 35kDa bands corresponding to isoforms of C2orf33 were observed across cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), DU 145 (Lane 2), tissue extracts of Mouse Heart (Lane 3) and Mouse Brain (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 (1:1000 dilution) and detected by Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 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 C2orf33 was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with C2orf33 Rabbit Polyclonal Antibody (Product # PA5-52765) at 5µg/mL dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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 composite image showing mitochondrial localization of C2orf33. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. .

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of C2orf33 in human gallbladder using a C2orf33 Polyclonal Antibody (Product # PA5-52765) shows weak cytoplasmic positivity in glandular cells.