Name: Anti-Protein CASP antibody [2A10]
SKU: ab54583
Rating: 4 (7 reviews)

Product name

Anti-Protein CASP antibody [2A10]
See all Protein CASP primary antibodies
Description

Mouse monoclonal [2A10] to Protein CASP
Host species

Mouse
Tested applications

Suitable for: IHC-P, ICC, WB, IHC-FoFr, ICC/IF, Sandwich ELISA, Flow Cytmore details
Species reactivity

Reacts with: Mouse, Human
Immunogen

Recombinant fragment (GST-tag) corresponding to Human Protein CASP aa 521-621. NP_001904.2
Sequence:
AENRLAQHTLQALQSELDSLRADNIKLFEKIKFLQSYPGRGSGSDDTELR YSSQYEERLDPFSSFSKRERQRKYLSLSPWDKATLSMGRLVLSNKMARTI

Database link: Q13948

Run BLAST with Run BLAST with
Positive control
- ICC/IF: HeLa cells. IHC-P: Human spleen tissue; human malignant lymphoma, diffuse large B tissue. Flow Cytometry: MCF7 cells. WB: Protein CASP transfected HEK-293T cell lysate. Recombinant Protein CASP protein.
General notes

This product was changed from ascites to tissue culture supernatant on 13^th Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.4
Constituent: PBS
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

2A10
Isotype

IgG1
Light chain type

kappa
Research areas

Compatible Secondaries
Immunohistochemistry kits
- Mouse on Mouse Polymer IHC Kit (ab127055)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Our Abpromise guarantee covers the use of ab54583 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
IHC-P		Use at an assay dependent concentration.
ICC		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration.
IHC-FoFr		Use at an assay dependent concentration. PubMed: 25355226
ICC/IF		Use at an assay dependent concentration.
Sandwich ELISA		Use at an assay dependent concentration.
Flow Cyt		Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Function

May be involved in intra-Golgi retrograde transport.
Sequence similarities

Belongs to the CASP family.
Cellular localization

Golgi apparatus membrane.
Target information above from: UniProt accession Q13948 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 1523 Human
- SwissProt: Q13948 Human
- SwissProt: P53564 Mouse
- Unigene: 191482 Human
Alternative names
- CASP_HUMAN antibody
- CUX1 antibody
- Protein CASP antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Protein CASP antibody [2A10] (ab54583)

Formalin-fixed, paraffin-embedded human spleen tissue stained for Protein CASP with ab54583 at 5 µg/ml in immunohistochemical analysis.

This image was generated using the ascites version of the product.
Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [2A10] (ab54583)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Protein CASP (green) using ab54583 at 10 µg/ml in ICC/IF.

This image was generated using the ascites version of the product.
Flow Cytometry - Anti-Protein CASP antibody [2A10] (ab54583)

Overlay histogram showing MCF7 (human breast adenocarcinoma cell line) cells stained with ab54583 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3 M glycine to block non-specific protein-protein interactions followed by the antibody (ab54583, 1 µg/1 x 10⁶ cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1 x 10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.
Western blot - Anti-Protein CASP antibody [2A10] (ab54583)

All lanes : Anti-Protein CASP antibody [2A10] (ab54583) at 5 µg/ml

Lane 1 : Protein CASP transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 2 : Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

This image was generated using the ascites version of the product.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Protein CASP antibody [2A10] (ab54583)

Formalin-fixed, paraffin-embedded human malignant lymphoma, diffuse large B tissue stained for Protein CASP with ab54583 at 5 µg/ml in immunohistochemical analysis.

This image was generated using the ascites version of the product.
Western blot - Anti-Protein CASP antibody [2A10] (ab54583)

Anti-Protein CASP antibody [2A10] (ab54583) at 5 µg/ml + Tagged recombinant Protein CASP protein

This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.

Expected MW 38 kDa.

This image was generated using the ascites version of the product.
Sandwich ELISA - Anti-Protein CASP antibody [2A10] (ab54583)

Detection limit of ab54583 is approximately 0.03 ng/ml as a capture antibody.

This image was generated using the ascites version of the product.

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

This product has been referenced in:

Zhang Z et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). IHC-Fr ; Mouse . Read more (PubMed: 30297870) »
Edri R et al. Analysing human neural stem cell ontogeny by consecutive isolation of Notch active neural progenitors. Nat Commun 6:6500 (2015). Other ; Human . Read more (PubMed: 25799239) »

See all 10 Publications for this product

Publishing research using ab54583? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (cortical neurons)

Permeabilization

Yes - 0.25% Triton X-100 in PBS

Specification

cortical neurons

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 06 2018

Application

Immunohistochemistry free floating

Sample

Mouse Tissue sections (P12 brain sections)

Specification

P12 brain sections

Dr. Bhavin Shah

Verified customer

Submitted Apr 17 2018

Abcam has not validated the combination of species/application used in this Abreview.

Application

Immunohistochemistry (Frozen sections)

Sample

Rabbit Tissue sections (brain)

Permeabilization

Yes - 0.3% triton

Specification

brain

Blocking step

Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Mar 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry

Sample

Human Cultured Cells (HeLa)

Permeabilization

Yes - 0.3% Triton-X-100 in PBS

Specification

HeLa

Blocking step

10% normal serum, 1% BSA in PBST as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted May 02 2013

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Sample

Rat Tissue sections (Brain)

Antigen retrieval step

None

Permeabilization

Specification

Brain

Fixative

Paraformaldehyde

Dr. Sophie Pezet

Verified customer

Submitted Jun 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (brain, placenta)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA, pH9, 20 min, 100C

Permeabilization

Specification

brain, placenta

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Nov 09 2011

Question

Dear Sir or Madam, I am interested in Anti-CUTL1 antibody and have some questions about it. There are 5 types anti-CUTL1 antibodies in the webpage. Anti-CUTL1 antibody (ab73885) looks better according to WB. What is the antibody's charaterization? Is it manufactured in-house or from outside labs? According to the reference, one of the antibodies is from xxxxxxxx group. Is ab73885 the right one? Many Thanks.

Abcam community

Verified customer

Asked on Oct 18 2011

Answer

Thank you for contacting us and sorry for the delay in getting back to you. We have a number of collaborators which produce antibodies for us as well as in house production of antibodies. The 5 antibodies that we have against CUTL1 have all been sourced through collaborators. All of the antibodies have been characterised through Western blotting and ab54583 and ab11584 has also been used in IHC. The diffferences observed in the western blots may be due to the antibodies being produced against different isoforms of CULT1. Whilst ab73885, ab85994 and ab98273 have been raised against immunogen sequences from CULT1 isoform 1 (SwissProt P39880) and would be expected to react with all CULT1 isoforms, ab115854 and ab54583 have both been raised against an immunogen sequence taken from CULT1 isoform 4 (SwissProt Q13948). These would therefore be expected to recognise isoform 4 and isoform 8 of CULT1 only (expected at around 77 kDa). I'm sorry if this information may not have been made explicit on the datasheet. Unfortunately I have no knowledge of any of the antibodies being produced by the group mentioned. You have not clarified the application or type of sample intended with the antibody but ab73885 would be a good choice if intending to perform Western blotting or ELISA with human CUTL1. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Oct 18 2011

Application

Western blot

Sample

Human Cell lysate - whole cell (MDA-MB-231, HT-1080, NIH3T3, D283Med, Daoy)

Gel Running Conditions

Reduced Denaturing (8%gel)

Loading amount

500000 cells

Specification

MDA-MB-231, HT-1080, NIH3T3, D283Med, Daoy

Blocking step

3% non fat milk/ 2% BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Ms. Sabine Topka

Verified customer

Submitted Oct 07 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-Protein CASP antibody [2A10] (ab54583)

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References

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Protein CASP antibody [2A10]

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Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Protein CASP antibody [2A10]

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