Anti-CBS antibody (ab96252)

Submit a review Q&A (1)References (4)

Overview

  • Product name

    Anti-CBS antibody
    See all CBS primary antibodies

  • Description

    Rabbit polyclonal to CBS

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Human, Zebrafish
    Predicted to work with: Mouse, Rat, Rabbit, Cow

  • Immunogen

    Recombinant fragment corresponding to a region within amino acids 39-221 of Human CBS (AAH00440).

  • Positive control

    • Molt-4 and Raji whole cell lysates; OVCAR3 xenograft; 293T, H1299 and HeLa whole cell lysate (ab150035).

Properties

Applications

Our Abpromise guarantee covers the use of ab96252 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
WB 1/500 - 1/3000. Predicted molecular weight: 61 kDa.
IHC-P 1/100 - 1/500.

Target

  • Function

    Only known pyridoxal phosphate-dependent enzyme that contains heme. Important regulator of hydrogen sulfide, especially in the brain, utilizing cysteine instead of serine to catalyze the formation of hydrogen sulfide. Hydrogen sulfide is a gastratransmitter with signaling and cytoprotective effects such as acting as a neuromodulator in the brain to protect neurons against hypoxic injury.

  • Tissue specificity

    In the adult strongly expressed in liver and pancreas, some expression in heart and brain, weak expression in lung and kidney. In the fetus, expressed in brain, liver and kidney.

  • Pathway

    Amino-acid biosynthesis; L-cysteine biosynthesis; L-cysteine from L-homocysteine and L-serine: step 1/2.

  • Involvement in disease

    Defects in CBS are the cause of cystathionine beta-synthase deficiency (CBSD) [MIM:236200]. CBSD is an enzymatic deficiency resulting in altered sulfur metabolism and homocystinuria. The clinical features of untreated homocystinuria due to CBS deficiency include myopia, ectopia lentis, mental retardation, skeletal anomalies resembling Marfan syndrome, and thromboembolic events. Light skin and hair can also be present. Biochemical features include increased urinary homocystine and methionine.

  • Sequence similarities

    Belongs to the cysteine synthase/cystathionine beta-synthase family.
    Contains 1 CBS domain.

  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AI047524 antibody
    • AI303044 antibody
    • Beta thionase antibody
    • Beta-thionase antibody
    • Cbs antibody
    • Cbs cystathionine beta-synthase antibody
    • CBS_HUMAN antibody
    • Cystathionine beta synthase antibody
    • Cystathionine beta-synthase antibody
    • EC 4.2.1.22 antibody
    • HIP 4 antibody
    • HIP4 antibody
    • Methylcysteine synthase antibody
    • MGC18856 antibody
    • MGC18895 antibody
    • MGC37300 antibody
    • OTTHUMP00000109416 antibody
    • OTTHUMP00000109418 antibody
    • Serine sulfhydrase antibody
    see all

Images

  • Western blot - Anti-CBS antibody (ab96252)
    Western blot - Anti-CBS antibody (ab96252)
    All lanes : Anti-CBS antibody (ab96252) at 1/5000 dilution

    Lane 1 : WT HeLa cell extract
    Lane 2 : CBS Knockout HeLa cell extracts

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 61 kDa

  • Western blot - Anti-CBS antibody (ab96252)
    Western blot - Anti-CBS antibody (ab96252)
    All lanes : Anti-CBS antibody (ab96252) at 1/5000 dilution

    Lane 1 : Molt-4 whole cell lysate
    Lane 2 : Raji whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 61 kDa



    7.5% SDS PAGE
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBS antibody (ab96252)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBS antibody (ab96252)
    Immunohistochemical analysis of paraffin-embedded OVCAR3 xenograft using ab96252 at 1/500 dilution.
  • Immunocytochemistry/ Immunofluorescence - Anti-CBS antibody (ab96252)
    Immunocytochemistry/ Immunofluorescence - Anti-CBS antibody (ab96252)
    ICC/IF image of ab96252 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab96252, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Western blot - Anti-CBS antibody (ab96252)
    Western blot - Anti-CBS antibody (ab96252)
    All lanes : Anti-CBS antibody (ab96252) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 20 µg
    Lane 4 : Zebrafish liver homogenate at 20 µg
    Lane 5 : Human epithelial carcinoma cell line at 20 µg

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 61 kDa
    Observed band size: 72 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes

References

This product has been referenced in:

  • Chen Y  et al. O-GlcNAcylated c-Jun antagonizes ferroptosis via inhibiting GSH synthesis in liver cancer. Cell Signal 63:109384 (2019). Read more (PubMed: 31394193) »
  • Song W  et al. Edoxaban improves venous thrombosis via increasing hydrogen sulfide and homocysteine in rat model. Mol Med Rep 16:7706-7714 (2017). Read more (PubMed: 28944880) »
See all 4 Publications for this product

Publishing research using ab96252? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question

DESCRIPTION OF THE PROBLEM One very strong band of wrong size (~35 kDa), one weak band of approximately correct size (~60 kDa) SAMPLE Cell lysate for MIN6 and INS-1E cell lines PRIMARY ANTIBODY polyclonal rabbit, abcam96252, dilution 1:1000 incubation 4-8 oC overnight DETECTION METHOD ECL Plus ANTIBODY STORAGE CONDITIONS aliquoted in 5 microliter quantities and frozen at -20 oC thawed at RT SAMPLE PREPARATION RIPA lysis buffer lysates stored at -80 oC and later thawed on ice Complete, Mini protease inhibitor added to lysis buffer WB samples prepared 1:3 ratio of 4x Laemmli reducing buffer to cell lysate heated at 92 oC for 5 min AMOUNT OF PROTEIN LOADED 15 micrograms ELECTROPHORESIS/GEL CONDITIONS Novex NuPAGE 4-12% Bis Tris gel TRANSFER AND BLOCKING CONDITIONS semi dry transfer buffer (48 mM Trisbase, 39 mM Gly, 1.3 mM SDS, 20% MeOH) blocking in 5% milk powder SECONDARY ANTIBODY goat anti rabbit HRP incubation RT 1 hr HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. -I would really appreciate if you could send us an image from the membrane to help us better understand the problem, but, regarding the description of the problem provided, I would say there might probably have been some kind of degradation of the sample, giving as a result a lower (half of the target MW) band. In that case, I would suggest optimising the sample preparation in order to avoid sample degradation. Please make sure phosphatase inhibitors are also used. -I would recommend loading a higher quantity of sample per lane; maybe 20ug would give better results. -Another suggestion I’d propose is increasing the primary antibody concentration to, for instance, 1/500, to obtain a stronger band at the expected molecular weight. -Running a no primary control would help dismiss any secondary antibody failure. - I would also appreciate if you could also provide details of your order (purchase order number, date of purchase, antibody lot number). I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact us again.

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