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CRALBP Monoclonal Antibody (B2)

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View details.

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Catalog # MA1-813

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CRALBP Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunostaining of CRALBP in frozen mouse retina sections using Product # MA1-813.

CRALBP Antibody — Fig 6 BMP/Wnt signalling is required for MET. A. Immunocytochemistry for PMEL17 where cells are seeded at either low density (16000 cells/cm 2 ) in the presence or absence of BMP4/7 (top left) or at high density (25000 cells/cm 2 ) in the presence or absence of Wnt5B (bottom left) and cultured for a period of 14 days. Also shown is the expression of BEST1 under the same conditions (top and bottom right). ACTB and B2M are used as housekeeping genes. Bars represent Mean + SD (n = 3). B. Quantification of immunocytochemistry for % CRALBP at Day 21 where cells are either treated with media alone (Control) or media supplemented with 10muM LDN-193189 added at Day 2,4,6,8,11,14 or 18. * indicates significant difference between control and compound treatment (One way ANOVA with Dunnett's multiple comparisons). C. Quantification of immunocytochemistry for % CRALBP at Day 28 where cells are either treated with media alone (Control) or media supplemented with 10muM WAY-262611 added at Day 2,7,14 or 21. * indicates significant difference between control and compound treatment (One way ANOVA with Dunnett's multiple comparisons). D. qPCR based measurement of BMP7 and Wnt5B transcript expression at Day 10 post siFOXM1 transfection (relative to transfection with non-targeting siRNA used as a control). GAPDH , HPRT1 and IPO8 were used as housekeeping genes. Bars represent Mean + SD (n = 3). P<0.05 (Student's t-test). {TM}

CRALBP Antibody in Western Blot (WB) — Figure 1. Immunophenotypic characterization of mouse MG cell line RG17. (A) Phase contrast images and immunofluorescence staining of primary MG cells and passage 20 RG17 cells. RG17 demonstrate typical MG morphology in culture and express many well-characterized MG markers. Scale bar, 50 um and applies to all immunofluorescence images. (B) Expression of characteristic MG genes in primary MG and RG17 cells (passages 20-25). The geometric means and geometric standard deviations ( N = 3 independent cultures) are graphed. An unpaired t -test was applied with a P value of <=0.05 considered statistically significant. (C) Western blot detection of MG markers from primary MG and passage 20 RG17 cells. (D) The doubling rate of RG17 cells does not vary significantly between passages 5 and 20. Data points are mean +- standard deviation.

CRALBP Antibody in Western Blot, Immunohistochemistry (WB, IHC) — Figure 3 Visual cycle proteins were abundantly expressed in the RPE flatmounts during the first 3 days in culture, but gradually decreased over time. RPE flatmounts obtained from 2- and 9-month-old wild-type C57BL/6J mice were cultured in complete medium for up to 7 days. RPE flatmounts treated with H 2 O 2 were cultured for 16 h. ( A ) Protein lysates were prepared from the RPE flatmounts and subjected to western blotting analysis to detect the levels of visual cycle proteins RPE65 and CRALBP. Statistical analysis was performed using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n = 3. ( B ) RPE65 and CRALBP protein levels were also measured by immunofluorescence. Data from both the western blotting and immunofluorescence showed the gradual reduction in the two proteins in RPE flatmounts over time. Scale bar = 20 mum.

CRALBP Antibody in Immunohistochemistry (IHC) — Fig. 6 Development of NR2E1 MiniPromoters for Muller glia cells. A Bioinformatic design of first-generation Ple264, cutdown to produce second-generation Ple315, and redesign to produce second-generation Ple316. ENCODE segments that potentially regulate the expression of the NR2E1 gene are highlighted horizontally. Vertically highlighted genomic regions correspond to their color-matched segments included in the MiniPromoter designs and are numbered as promoter(s) (P) and regulatory region(s) (RR). B Postnatal day 4 intravenous injection of Ple316-EmGFP, harvested 4 weeks later, led to robust expression with cell bodies in the inner nuclear layer (INL) and processes that span the retina. Scale bar, 100 um. (For ubiquitous smCBA promoter see Fig. S1A .) C Adult intravitreal injection of Ple316-EmGFP, harvested 4 weeks later, was quantified for epifluorescence intensity in CRALBP-positive cells, and showed no significant difference when compared to smCBA. (For smCBA see Fig. S1D ). D Adult intravitreal injection of Ple316-EmGFP, harvested 4 weeks later, led to robust expression in the Muller glia cells, as indicated by co-staining with Muller glia cell marker CRALBP. Scale bar, 20 um. (For smCBA see Fig. S1D .) EmGFP emerald green fluorescent protein, GCL ganglion cell layer, IPL inner plexiform layer, N number of cells counted, ONL outer nuclear layer, OPL outer plexiform layer, TF transcription factor, TSS transcription start site. Green, anti-GFP; blue, Hoechst; red, anti-CRAL

CRALBP Antibody in Immunocytochemistry (ICC/IF) — Stage 3: Generation of a homogenous and functional retinal pigment epithelium (RPE) population. (A): Two days' postseeding, SHEF1 human embryonic stem cells were treated with Induction Medium 1 for 4 days, followed by Induction Medium 2 for 3 days. At day 9, cells were replated in Induction Medium 3 for a period of 19 days. At day 28, cells were replated in Dulbecco's modified Eagle's medium KSR-XF and cultured for a period of 14 days. Representative images showing immunocytochemistry for indicated RPE markers are shown. Images are captured at x10 magnification. (B): Cells cultured as in panel A were further dissociated and replated on transwells and cultured for a period of 10 weeks. A representative image showing en face immunocytochemistry for ZO-1 is shown. Images are captured at x20 magnification. (C): Spent medium from transwells described in panel B was collected from the top and bottom chambers and quantified for vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) concentration. The ratio of [VEGF]:[PEDF] was quantified in media from the two compartments ( n = 3, +-SD). (D): Representative confocal images showing phagocytosis of fluorescent bead (red) by RPE. ZO-1 immunocytochemistry (green) shows the cell edge, and the presence of bead within the cell boundary indicates internalization by phagocytosis. Images are captured at x63 magnification. Abbreviations: PEDF, pigment epithelium-derived factor; VEGF, vascular endo

CRALBP Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Published figure using CRALBP monoclonal antibody (Product # MA1-813) in Western Blot and Immunocytochemistry

Product Details

TESTED APPLICATIONS	DILUTION
Flow Cytometry (Flow)	Assay-dependent
Immunohistochemistry (Frozen) (IHC (F))	1 µg/mL
Immunoprecipitation (IP)	Assay-dependent
Western Blot (WB)	0.02 µg/mL

PUBLISHED APPLICATIONS
Western Blot (WB)	See 5 publications below
Immunohistochemistry (IHC)	See 13 publications below
Immunohistochemistry (Paraffin) (IHC (P))	See 1 publications below
Immunocytochemistry (ICC/IF)	See 9 publications below
Miscellaneous PubMed (Misc)	See 1 publications below
Flow Cytometry (Flow)	See 1 publications below

Species reactivity	Bovine, Human, Mouse, Non-human primate, Rat
Published species	Human, Mouse, Rat
Host / Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	B2
Immunogen	Human recombinant CRALBP.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2178528

Product Specific Information

MA1-813 detects CRALBP protein in bovine, human, mouse, non-human primate and rat samples.

MA1-813 has successfully been used in Western blot, immunohistochemistry, immunoprecipitation, and immunofluorescence procedures. By Western blot, this antibody detects a 37 kDa protein representing human recombinant CRALBP.

The MA1-813 immunogen is purified human recombinant CRALBP.

Background/Target Information

Cellular retinaldehyde-binding protein (CRALBP) plays an important role in the regeneration of 11-cis-retinal for use in rod visual pigments such as opsin and rhodopsin. Once 11-cis-retinal is photoisomerized in the rod outer segment, it is converted to all-trans-retinal and further modified into all-trans-retinol. All-trans-retinol then diffuses into the retinal pigment epithelium (RPE) to be converted back to 11-cis-retinol and further oxidized into 11-cis-retinal (both by CRALBP). Genetic mutations involving CRALBP's lack of function have been linked to visual disease such as bothnia dystrophy, retinitis punctata albescens, retina pigmentosa, and Newfoundland rod-cone dystrophy. The presence of CRALBP serves as a marker for RPE and Muller glial cells of the retina.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

References (16)

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Bioinformatics

Protein Aliases: Cellular retinaldehyde-binding protein; cellular retinaldehyde-binding protein-1; Retinaldehyde-binding protein 1; retinaldehyde-binding protein precursor

Gene Aliases: 3110056M11Rik; CRALBP; RLBP1

UniProt ID: (Human) P12271, (Bovine) P10123, (Mouse) Q9Z275

Entrez Gene ID: (Human) 6017, (Bovine) 282038, (Mouse) 19771, (Rat) 293049

Molecular Function: Blue Arrow Icon transfer/carrier protein

Function(s)

retinoid metabolic process vitamin A metabolic process transport visual perception response to stimulus biological_process

Process(es)

transporter activity
11-cis retinal binding
retinol binding
molecular_function

Suggested Secondary Antibodies

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HRP Anti-Mouse Secondary Antibodies (234)

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CRALBP Monoclonal Antibody (B2)

Product Details

Product Specific Information

Background/Target Information

References (16)

Have you cited this antibody in a publication? so we can reference it in this datasheet.

Bioinformatics

Suggested Secondary Antibodies

Documents

Material safety data sheets (MSDS)

Performance Guarantee

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CRALBP Monoclonal Antibody (B2)

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Product Details

Product Specific Information

Background/Target Information

References (16)

Bioinformatics

Suggested Secondary Antibodies

Documents

Material safety data sheets (MSDS)

Performance Guarantee

We’re here to help

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