Anti-CD40 antibody (ab113701)
Key features and details
- Rabbit polyclonal to CD40
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-CD40 antibody
See all CD40 primary antibodies -
Description
Rabbit polyclonal to CD40 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: U-2 OS and Raji cell lysates; Human spleen and thymus tissue lysates. IHC-FFPE: Human spleen tissue.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab113701 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species IHC-P HumanWB HumanAll applications Macaque monkeyGorillaApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 31 kDa).IHC-P Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 31 kDa).IHC-P
Use a concentration of 5 µg/ml.Target
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Function
Receptor for TNFSF5/CD40LG. -
Tissue specificity
B-cells and in primary carcinomas. -
Involvement in disease
Defects in CD40 are the cause of hyper-IgM immunodeficiency syndrome type 3 (HIGM3) [MIM:606843]; also known as hyper-IgM syndrome 3. HIGM3 is an autosomal recessive disorder which includes an inability of B cells to undergo isotype switching, one of the final differentiation steps in the humoral immune system, an inability to mount an antibody-specific immune response, and a lack of germinal center formation. -
Sequence similarities
Contains 4 TNFR-Cys repeats. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 958 Human
- Omim: 109535 Human
- SwissProt: P25942 Human
- Unigene: 472860 Human
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Alternative names
- AI326936 antibody
- B cell associated molecule CD40 antibody
- B cell surface antigen CD40 antibody
see all
Images
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Western blot - Anti-CD40 antibody (ab113701)All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml
Lane 1 : Wild-type U-2 OS cell lysate
Lane 2 : CD40 knockout U-2 OS cell lysate
Lane 3 : Raji cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab113701 observed at 45 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab113701 was shown to react with CD40 in wild-type U-2 OS cells in Western blot. Loss of signal was observed when CD40 knockout sample was used. Wild-type and CD40 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab113701 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-CD40 antibody (ab113701)All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml
Lane 1 : Human spleen tissue lysate - total protein (ab29699)
Lane 2 : Human thymus tissue lysate - total protein (ab30146)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?
Additional bands at: 105 kDa, 41 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.
Exposure time: 10 minutes
CD40 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The predicted molecular weight of CD40 is 31 kDa (SwissProt), however we expect to observe a banding pattern around 43 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab113701)IHC image of CD40 staining in Human normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab113701, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab113701 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-CD40 antibody (ab113701)All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml
Lane 1 : Wild-type U-2 OS cell lysate
Lane 2 : CD40 knockout U-2 OS cell lysate
Lane 3 : Raji cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab113701 observed at 45 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab113701 was shown to react with CD40 in wild-type U-2 OS cells in Western blot. Loss of signal was observed when CD40 knockout sample was used. Wild-type and CD40 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab113701 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-CD40 antibody (ab113701)All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml
Lane 1 : Human spleen tissue lysate - total protein (ab29699)
Lane 2 : Human thymus tissue lysate - total protein (ab30146)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?
Additional bands at: 105 kDa, 41 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.
Exposure time: 10 minutes
CD40 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The predicted molecular weight of CD40 is 31 kDa (SwissProt), however we expect to observe a banding pattern around 43 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab113701)
IHC image of CD40 staining in Human normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab113701, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
