CHM Recombinant Rabbit Monoclonal Antibody [JE61-83]
Catalog# HA720064
CHM Recombinant Rabbit Monoclonal Antibody [JE61-83]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
CHM Recombinant Rabbit Monoclonal Antibody [JE61-83]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human CHM aa 36-85/653.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
Predicted band size: 73 kDa
Positive Control
A431 cell lysate, SiHa cell lysate, C2C12 cell lysate, C6 cell lysate, mouse colon tissue lysate, rat colon tissue lysate, human colon carcinoma tissue, human endometrium tissue, mouse brain tissue, mouse colon tissue, rat rectum tissue, SiHa, HeLa.
Conjugation
unconjugated
Clone Number
JE61-83
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:50
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IHC-P
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1:200-1:1,000
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FC
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1:500-1:1,000
Target
Function
This gene encodes component A of the RAB geranylgeranyl transferase holoenzyme. In the dimeric holoenzyme, this subunit binds unprenylated Rab GTPases and then presents them to the catalytic Rab GGTase subunit for the geranylgeranyl transfer reaction. Rab GTPases need to be geranylgeranyled on either one or two cysteine residues in their C-terminus to localize to the correct intracellular membrane. Mutations in this gene are a cause of choroideremia; also known as tapetochoroidal dystrophy (TCD). This X-linked disease is characterized by progressive dystrophy of the choroid, retinal pigment epithelium and retina. Alternatively spliced transcript variants have been found for this gene. Besides, a pre-formed complex consisting of CHM and the Rab GGTase dimer (RGGT or component B) can bind to and prenylate Rab proteins; this alternative pathway is proposed to be the predominant pathway for Rab protein geranylgeranylation.
Background References
1. Murro V. et. al. Optical Coherence Tomography Angiography (OCT-A) in Choroideremia (CHM) carriers. Ophthalmic Genet. 2020 Apr
2. Imani S. et. al. Molecular genetics characterization and homology modeling of the CHM gene mutation: A study on its association with choroideremia. Mutat Res. 2018 Jan-Mar
Subcellular Location
Cytoplasm, cytosol.
Synonyms
CHM antibody
Chm antibody
Choroideraemia protein antibody
Choroideremia antibody
DXS540 antibody
FLJ38564 antibody
GGTA antibody
HSD 32 antibody
MGC102710 antibody
Rab escort protein 1 antibody
ExpandImages
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Western blot analysis of CHM on different lysates with Rabbit anti-CHM antibody (HA720064) at 1/5,000 dilution.
Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: SiHa cell lysate (20 µg/Lane)
Lane 3: C2C12 cell lysate (20 µg/Lane)
Lane 4: C6 cell lysate (20 µg/Lane)
Lane 5: Mouse colon tissue lysate (40 µg/Lane)
Lane 6: Rat colon tissue lysate (40 µg/Lane)
Predicted band size: 73 kDa
Observed band size: 100 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720064) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-CHM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720064, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human endometrium tissue using anti-CHM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720064, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-CHM antibody (HA720064) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720064) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-CHM antibody (HA720064) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720064) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat rectum tissue using anti-CHM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720064, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of CHM was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (HA720064, 1ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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Immunocytochemistry analysis of HeLa cells labeling CHM with Rabbit anti-CHM antibody (HA720064) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CHM antibody (HA720064) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) were used as the secondary antibody at 1/1,000 dilution.
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