HA720064

CHM Recombinant Rabbit Monoclonal Antibody [JE61-83]

cat.: HA720064

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE61-83

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	73 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human CHM aa 36-85/653.
Positive control:	Mouse colon tissue lysates, human colon carcinoma tissue, human endometrium tissue, rat rectum tissue, SiHa, Hela.
Subcellular location:	Cytosol.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:2,000 1:50 1:50-1:200 1:500-1:1,000
Uniprot #:	SwissProt: P24386 Human \| Q9QXG2 Mouse \| P37727 Rat
Alternative names:	CHM Chm Choroideraemia protein Choroideremia DXS540 FLJ38564 GGTA HSD 32 MGC102710 Rab escort protein 1 Rab geranylgeranyltransferase component A Rab proteins geranylgeranyltransferase component A 1 RAE1_HUMAN REP 1 REP-1 REP1 TCD TCD protein

Images

	Fig1: Western blot analysis of CHM on mouse colon tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720064, 1/500) was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 73 kDa Observed band size: 105 kDa
	Fig2: 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.
	Fig3: 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.
	Fig4: 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.

Fig5: 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).

Fig6: 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.