Overview

  • Product name
  • Description
    Rabbit polyclonal to Annexin IV
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig
  • Immunogen

    A region within Synthetic peptide: GLGTDEDAII SVLAYRNTAQ RQEIRTAYKS TIGRDLIDDL KSELSGNFEQ , corresponding to N terminal amino acids 31-80 of Human Annexin IV

  • Positive control
    • WB: Placental lysate. IHC-P: Human intestine.

Properties

Applications

Our Abpromise guarantee covers the use of ab33009 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-Fr 1/100.
ELISA 1/1.5625e+006.
WB Use a concentration of 1.25 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 36 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use at an assay dependent concentration.

Target

  • Function
    Calcium/phospholipid-binding protein which promotes membrane fusion and is involved in exocytosis.
  • Sequence similarities
    Belongs to the annexin family.
    Contains 4 annexin repeats.
  • Domain
    A pair of annexin repeats may form one binding site for calcium and phospholipid.
  • Information by UniProt
  • Database links
  • Alternative names
    • 35 beta calcimedin antibody
    • 35-beta calcimedin antibody
    • AIV antibody
    • Annexin 4 antibody
    • Annexin A4 antibody
    • Annexin IV (placental anticoagulant protein II) antibody
    • Annexin IV antibody
    • Annexin IV placental anticoagulant protein II antibody
    • Annexin-4 antibody
    • AnnexinA4 antibody
    • AnnexinIV antibody
    • ANX 4 antibody
    • ANX A4 antibody
    • ANX4 antibody
    • ANXA 4 antibody
    • ANXA4 antibody
    • ANXA4 protein antibody
    • ANXA4_HUMAN antibody
    • Carbohydrate binding protein P33/P41 antibody
    • Carbohydrate-binding protein p33/p41 antibody
    • Chromobindin 4 antibody
    • Chromobindin-4 antibody
    • Chromobindin4 antibody
    • DKFZp686H02120 antibody
    • Endonexin I antibody
    • EndonexinI antibody
    • Lipocortin IV antibody
    • LipocortinIV antibody
    • MGC75105 antibody
    • OTTHUMP00000160052 antibody
    • OTTHUMP00000202207 antibody
    • P32.5 antibody
    • P33/41 antibody
    • PAP II antibody
    • PAP-II antibody
    • PAPII antibody
    • PIG 28 antibody
    • PIG28 antibody
    • Placental anticoagulant protein II antibody
    • PP4 X antibody
    • PP4-X antibody
    • PP4X antibody
    • Proliferation inducing gene 28 antibody
    • Proliferation inducing protein 28 antibody
    • Protein II antibody
    • Xanx-4 antibody
    • ZAP 36 antibody
    • ZAP36 antibody
    see all

Images

  • Anti-Annexin IV antibody (ab33009) at 1.25 µg/ml + Placental lysate

    Secondary
    HRP conjugated anti Rabbit IgG 1/50000 - 1/100000

    Predicted band size: 36 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human intestine tissue labelled at Annexin IV with ab33009 at 4-8µg/ml. Positively labelled cells (indicated by arrows) are epithelial cells of intestinal villi. Magnification: 400X.

References

This product has been referenced in:
  • Choi CH  et al. Prognostic significance of annexin A2 and annexin A4 expression in patients with cervical cancer. BMC Cancer 16:448 (2016). Read more (PubMed: 27402115) »
  • Chen W  et al. Overexpression of annexin A4 indicates poor prognosis and promotes tumor metastasis of hepatocellular carcinoma. Tumour Biol 37:9343-55 (2016). Read more (PubMed: 26779633) »
See all 7 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Sorry for the delay in getting back to you.

I can now provide further advice for the customer. Whilst the antibody anti-Annexin IV (ab33009) will not be able to detect the fusion construct the customer has been expressing (with ANXA4 reidues 1-31), the antibody ab65846 would be suitable. This has been raised against a peptide taken from within the residues 1-30 on human Annexin IV and should therefore be able to detect the region of the protein your custoemr has been expressing.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact ua again.

Read More

Answer

Thank you very much for this information and I am sorry for the delay in getting back to you. I have been away from the office.

With this information provided, I now think I know why the experiment has not been working for the customer. The antibody anti-Annexin IV antibody (ab33009) which they have been using has been raised against the residues 31-80 of Human Annexin IV. This is therefore the region of the protein that the antibody will detect. It will not be able to detect the residues 1-31 which the customer has used in their fusion construct.

I am currently seeking information regarding a different antibody raised against Annexin IV to see if this would be more appropriate for your customer to use. I will get back to you with this information as soon as I have it.

I am sorry for the delay and any inconvenience this may cause you.

Read More

Answer

Thank you very much for sharing that information with me. It has been very helpful.

In order to ascertain if the antibody Anti-Annexin IV antibody (ab33009) is likely to detect the protein you have been expressing, could you please let me know exactly which protein of the human protein has been cloned into the pET30b vector?

You mentioned that only a partial length sequence of Anxa4 was used. Exactlywhat sequence was this? It may be that our antibody (ab33009) is directed towards a different part of the protein to the one included in your construct.

I look forward to receiving your reply.

Read More

Question

Product code: 33009
Lot number: GR16609-6

Inquiry: 1. Order details: - Batch number: lot:GR16609-6 - Po: 1096734 - Abcam product code: ab33009 - Antibody storage conditions (temperature/reconstitution etc) :-20℃ 2. Please describe the problem (high background, wrong band size, more bands, no band etc). Wrong band size 3. On what material are you testing the antibody in WB? - Species: human - What’s cell line or tissue: - Cell extract or Nuclear extract: - Purified protein or Recombinant protein: Recombinant protein 3. The lysate - How much protein was loaded: - What lysis buffer was used: - What protease inhibitors were used: - What loading buffer was used: - Phosphatase inhibitors - Did you heat the samples: temperature and time: 4. Electrophoresis/Gel conditions/ Transfer conditions - Reducing or non reducing gel: Reducing gel - Reducing agent: - Gel percentage : 10% - Transfer conditions: (Type of membrane, Protein transfer verified): PVDF 5. Blocking conditions - Buffer: TTBS - Blocking agent: milk, BSA, serum, what percentage: 5% milk in TTBS - Incubation time:1 hr - Incubation temperature: Room temperature 6. Primary Antibody - Species:Rabbit v - Reacts against: human - At what dilution(s) have you tested this antibody:1000x - What dilution buffer was used: 5% milk in TTBS - Incubation time: 1 hr - Incubation temperature: Room temperature - What washing steps were done: 10min in TTBS, 3 times 7. Secondary Antibody - Species:Donkey - Reacts against: Rabbit - At what dilution(s) have you tested this antibody: 10000x - Incubation time: 1 hr - Wash steps: 10min in TTBS, 3 times - Fluorochrome or enzyme conjugate: HRP - Do you know whether the problems you are experiencing come from the secondary? NO 8. Detection method ECl, ECl+, other detection method: ECL+ 9. Did you apply positive and negative controls along with the samples? Please specify. Recombinant protein 10. Optimization attempts * How many times have you tried the Western? once * Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no * Do you obtain the same results every time e.g. are background bands always in the same place? * What steps have you altered? no

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibodyhas not been providing you with the expected results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would like to clarify a few points in order to understand what may be contributing to the problems encountered.

1. You have mentioned that a recombinant protein was used with the antibody. How was this obtained, was it purchased or expressed in a cell line? What was the sequence of this expressed protein.

2. Did you stain the SDS gel or membrane to ascertain the size of the recombinant protein before performing the Western blot? Could you share some images of this staining with me?

3. How was the sample prepared prior to loading it on the gel? How much protein was loaded? Was any protease inhibitors used in the lysis/loading buffer? Was the sample denatured by heating to 95 degrees for >5 minutes?

Once Ihave receivedthis information Iwill hopefully understand more fully what may be contributing totheproblems encountered.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Small intestinal epithelial cells)
Specification
Small intestinal epithelial cells
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C

Dr. Abeer Hammoudi

Verified customer

Submitted May 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (small intestine -epithelial cells extract)
Loading amount
20 µg
Specification
small intestine -epithelial cells extract
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Abeer Hammoudi

Verified customer

Submitted May 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (cartilage)
Specification
cartilage
Fixative
Methanol
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5

Abcam user community

Verified customer

Submitted Aug 08 2006

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

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