PA5-29300

antibody from Invitrogen Antibodies

Targeting: BCKDHA MSU, OVD1A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-29300

Provider

Invitrogen Antibodies

Product name

BCKDHA Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: A549, HeLa, HepG2.

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using BCKDHA Polyclonal Antibody (Product # PA5-29300). Sample (30 µg of whole cell lysate). Lane A: HepG2. 10% SDS PAGE. BCKDHA Polyclonal Antibody (Product # PA5-29300) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of BCKDHA was achieved by transfecting Hep G2 cells with BCKDHA specific siRNAs (Silencer® select Product # s1910, s1911). Western blot analysis (Fig. a) was performed whole cell extracts from the Hep G2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with BCKDHA Polyclonal Antibody (Product # PA5-29300, 1:1000 dilution) and Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 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 BCKDHA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-BCKDHA Rabbit Polyclonal Antibody (Product # PA5-29300) and a 50kDa band corresponding to BCKDHA was observed across all cell lines tested. Whole cell extracts (30 µg lysate) of HT-29 (Lane 1) and Hep G2 (Lane 2) 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 chemiluminescence 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

Immunocytochemistry-Immunofluorescence analysis of BCKDHA was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: BCKDHA Polyclonal Antibody (Product # PA5-29300) diluted at 1:400. Blue: Hoechst 33342 staining.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using BCKDHA (Product # PA5-29300) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.