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[KD Validated] CHCHD4 Rabbit mAb (A27252)

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 [KD Validated] CHCHD4 Rabbit mAb (A27252) at 1:13000 dilution incubated at room temperature for 1.5 hours.
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: 10s.

Western blot analysis of lysates from wild type (WT) and CHCHD4 knockdown (KD) 293T cells using [KD Validated] CHCHD4 Rabbit mAb (A27252) at 1:13000 dilution incubated at room temperature for 1.5 hours.
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: 10s.

ABclonal:Immunohistochemistry - [KD Validated] CHCHD4 Rabbit mAb (A27252)

Immunohistochemistry analysis of paraffin-embedded Human breast cancer tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human kidney tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat lung tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat testis tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KD Validated] CHCHD4 Rabbit mAb (A27252)

Confocal imaging of A-431 cells using [KD Validated] CHCHD4 Rabbit mAb (A27252, 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 Hep G2 cells using [KD Validated] CHCHD4 Rabbit mAb (A27252, 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 paraffin-embedded Human kidney tissue using [KD Validated] CHCHD4 Rabbit mAb (A27252, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.

Overview

Product name	[KD Validated] CHCHD4 Rabbit mAb
Catalog No.	A27252
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC72750

Background

CHCHD4, a component of human mitochondria, belongs to a protein family whose members share 6 highly conserved cysteine residues constituting a -CXC-CX(9)C-CX(9)C- motif in the C terminus (Hofmann et al., 2005 [PubMed 16185709]).

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-105 of human CHCHD4 (NP_001091972.1).
Sequence	MSYCRQEGKDRIIFVTKEDHETPSSAELVADDPNDPYEEHGLILPNGNINWNCPCLGGMASGPCGEQFKSAFSCFHYSTEEIKGSDCVDQFRAMQECMQKYPDLY
Gene ID
Swiss Prot
Synonyms	MIA40; TIMM40
Calculated MW	16kDa
Observed MW	22kDa

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 1:13000 - 1:78000 IHC-P 1:600 - 1:3000 IF/ICC 1:50 - 1:200 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 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	A-549, Mouse brain, Mouse heart, Rat testis, 293T
Cellular location	Mitochondrion intermembrane space.