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HADHA Rabbit PolymAb® (A24055PM)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using [KO Validated] HADHA Rabbit PolymAb® (A24055PM) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.

Western blot analysis of lysates from wild type (WT) and HADHA knockout (KO) 293T cells using [KO Validated] HADHA Rabbit PolymAb® (A24055PM) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.

ABclonal:Immunohistochemistry - HADHA Rabbit PolymAb® (A24055PM)

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using HADHA Rabbit PolymAb® (A24055PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human liver tissue using HADHA Rabbit PolymAb® (A24055PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human thyroid cancer tissue using HADHA Rabbit PolymAb® (A24055PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - HADHA Rabbit PolymAb® (A24055PM)

Confocal imaging of HeLa cells using HADHA Rabbit PolymAb® (A24055PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500)(Red).The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green).DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of PC-12 cells using HADHA Rabbit PolymAb® (A24055PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500)(Red).The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green).DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - HADHA Rabbit PolymAb® (A24055PM)

Immunoprecipitation of HADHA in 300 µg extracts from 293F cells using 3 µg HADHA Rabbit PolymAb® (A24055PM). Western blot analysis was performed using HADHA Rabbit PolymAb® (A24055PM) at 1:1000 dilution.

Overview

Product name	HADHA Rabbit PolymAb®
Catalog No.	A24055PM
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

This gene encodes the alpha subunit of the mitochondrial trifunctional protein, which catalyzes the last three steps of mitochondrial beta-oxidation of long chain fatty acids. The mitochondrial membrane-bound heterocomplex is composed of four alpha and four beta subunits, with the alpha subunit catalyzing the 3-hydroxyacyl-CoA dehydrogenase and enoyl-CoA hydratase activities. Mutations in this gene result in trifunctional protein deficiency or LCHAD deficiency. The genes of the alpha and beta subunits of the mitochondrial trifunctional protein are located adjacent to each other in the human genome in a head-to-head orientation.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 545-763 of human HADHA (NP_000173.2).
Sequence	FYTTRCLAPMMSEVIRILQEGVDPKKLDSLTTSFGFPVGAATLVDEVGVDVAKHVAEDLGKVFGERFGGGNPELLTQMVSKGFLGRKSGKGFYIYQEGVKRKDLNSDMDSILASLKLPPKSEVSSDEDIQFRLVTRFVNEAVMCLQEGILATPAEGDIGAVFGLGFPPCLGGPFRFVDLYGAQKIVDRLKKYEAAYGKQFTPCQLLADHANSPNKKFYQ
Gene ID
Swiss Prot
Synonyms	GBP; ECHA; HADH; LCEH; MTPA; LCHAD; TP-ALPHA
Calculated MW	83kDa
Observed MW

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:1000 IHC-P 1:50 - 1:200 IF/ICC 1:50 - 1:200 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Immunoprecipitation
Positive samples	293T, NIH/3T3, Mouse kidney, Rat kidney
Cellular location	Mitochondrion.